Methods for inducing immune responses

Abstract

Provided are methods for inducing immune responses against pathogens in a mammal. The methods comprise administering a single cycle adenovirus (SC-Ad) comprising a nucleic acid sequence encoding an immunogen of said pathogen to said mammal. The SC-Ad is administered by inhalation and infects a cell of said mammal, thereby causing expression of said immunogen leading to the induction of said immune response. New infectious virions are not produced. The methods may be of particular utility in vaccination for respiratory pathogens. Corresponding medical uses of the SC-Ads are also provided.

Description

[0001] METHODS FOR INDUCING IMMUNE RESPONSES

[0002] FIELD OF INVENTION

[0003] The invention relates to a method for reducing an immune responseto a pathogen in a mammal, in which a single cycle adenovirus comprising a nucleic acid encoding an immunogen of said pathogen is administered to said mammal by inhalation. The invention also relates to pharmaceutical compositions suitable for use in such methods, and to corresponding medical uses of single cycle adenoviruses.

[0004] BACKGROUND

[0005] Respiratory pathogens are microorganisms that cause infections primarily affecting the respiratory tract, which includes the upper respiratory tract (nose, throat, sinuses) and the lower respiratory tract (bronchi, lungs). These pathogens can include viruses, bacteria, fungi, and other infectious agents. Respiratory infections are among the most common infectious diseases worldwide and can range from mild illnesses, such as the common cold, to severe respiratory syndromes, such as pneumonia and acute respiratory distress syndrome (ARDS). A prominent example is COVID-19. This infectious disease causedby a coronavirus known as COVID-19 was first reported to the World Health Organization (WHO) Country Office in China on Decembers 1, 2019. As of June 3, 2020, approximately 6,287,771 confirmed cases of COVID- 19, including 379,941 deaths, have been reported to the WHO (covidl9.who.int / ).

[0006] Despite progress in vaccination and public health measures, treatments for respiratory pathogens remain indispensable for various reasons. Firstly, the emergence of new variants with altered characteristics necessitates ongoing development of treatments to effectively respond to evolving threats. Secondly, breakthrough infections can occur, especially in vulnerable populations or areas with incomplete vaccine coverage, highlighting the need for treatments to manage cases and reduce disease severity. Thirdly, certain populations, such as the elderly or immunocompromised, may still be at risk of severe illness, underscoring the importance of treatments in providing effective care. Moreover, the development of antiviral resistance and the occurrence of co-infections further emphasize the necessity of continuous research and development of new treatments. Indeed, there remains a need to identify new therapies for the treatment of respiratory pathogens, especially those which can be directly target to the respiratory mucosa such as treatments administered via inhalation.

[0007] Having the ability to produce immune responses effectively against viral and / or bacterial pathogens (e.g., a coronavirus) in mammals (e.g., humans) can improve survival and minimize the impact of an infection such as a respiratory infection.

[0008] One recent advancement is the use of adenovirus vectors. In traditional adenovirus vectors, the virus is capable of multiple rounds of replication, which can potentially cause safety concerns. Replication-competent Ad vectors have the potential to cause adverse effects, including excessive immune responses or inflammation, especially in immunocompromised individuals or those with pre-existing respiratory conditions. Uncontrolled viral replication may lead to systemic dissemination and severe complications.

[0009] The inventors have identified methods of treatment and medical uses that may solve one or more of the problems discussed above.

[0010] SUMMARY OF THE INVENTION

[0011] A first aspect of the invention provides a method for inducing an immune response against a pathogen in a mammal, the method comprising administering a single cycle adenovirus (SC- Ad) comprising a nucleic acid sequence encoding an immunogen of said pathogen to said mammal, wherein said SC-Ad is administered by inhalation and infects a cell of said mammal, thereby causing expression of said immunogen leading to the induction of said immune response, without production of new infectious virions.

[0012] In a suitable embodiment said SC-Ad is formulated as a liquid formulation.

[0013] In a suitable embodiment the liquid formulation comprises NaCl, histidine, MgCl2, EDTA, polysorbate 80, sucrose, and ethanol.

[0014] In a suitable embodiment said SC-Ad is formulated as a solid powder.

[0015] In a suitable embodiment said SC-Ad is administered in an aerosol. In a suitable embodiment said SC-Ad is administered in a therapeutically effective amount of at least 1 x 1010SC-Ad viral particles.

[0016] In a suitable embodiment said SC-Ad is administered in a therapeutically effective amount of between 1 x 1010and 3 x 1011SC-Ad viral particles.

[0017] In a suitable embodiment the administration of said SC-Ad by inhalation is provided as a vaccine boostto a mammal that has received a previous vaccination to the pathogen before said subsequent administration of said SC-Ad by inhalation.

[0018] In a suitable embodiment the administration of said SC-Ad by inhalation is provided as a vaccine boost to a mammal that has received a previous vaccination to the pathogen at least 90 days before said subsequent administration of said SC-Ad by inhalation.

[0019] In a suitable embodiment said prior vaccination did not comprise administration of said SC- Ad.

[0020] In a suitable embodiment said prior vaccine comprised administration of said SC-Ad.

[0021] In a suitable embodiment said pathogen is a respiratory pathogen.

[0022] In a suitable embodiment the immunogen of said respiratory pathogen is selected from the group consisting of: a SARS-CoV-2 immunogen, a SARS-CoV immunogen, a HCoV NL63 immunogen, a HKU1 immunogen, a MERS-CoV immunogen, an influenza immunogen, a respiratory syncytial virus (RSV) immunogen, a metapneumovirus (MPV) immunogen, a rhinovirus immunogen, a bocavirus immunogen, a parainfluenza virus (PIV) immunogen, a Streptococcus pneumoniae immunogen, a Bordetella pertussis immunogen, a Haemophilus influenzae immunogen, a Mycobacterium tuberculosis immunogen, a Klebsiella pneumonia immunogen, and an immunogen of a fungal respiratory pathogen.

[0023] In a suitable embodiment said pathogen is a coronavirus and the immunogen is a coronavirus immunogen comprisinga coronavirus Spike polypeptide, or an immunogenic fragment thereof In a suitable embodiment said pathogen is a Marburgvirus (such as Marburg virus or Ravn virus), or an ebolavirus.

[0024] In a suitable embodiment a mucosal immune response is induced.

[0025] In a suitable embodiment said mucosal immune response leads to an increase in the amount of serum IgG antibodies that bind to said immunogen.

[0026] In a suitable embodiment said mucosal immune response leads to an increase in the amount of IgA antibodies that bind to said immunogen.

[0027] In a suitable embodiment expression of said immunogen in said cell leads to an increase in the amount of serum IgA antibodies that bind to said immunogen

[0028] In a suitable embodiment expression of said immunogen in said cell leads to an increase in the amount of secretory IgA antibodies that bind to said immunogen

[0029] In a suitable embodiment said SC-Ad comprises a genomelacking at least a portion of a nucleic acid sequence that encodes an adenovirus polypeptide, and wherein said SC-Ad comprises said adenovirus polypeptide.

[0030] In a suitable embodiment said adenovirus polypeptide is selected from the group consisting of a fiber polypeptide, a V polypeptide, a hexon polypeptide, a penton base polypeptide, and a pllla polypeptide.

[0031] According to a second aspect of the invention, there is provided a single cycle adenovirus (SC- Ad) comprising a nucleic acid sequence encoding an immunogen of a pathogen for use in inducing an immune response against the pathogen in a mammal, wherein the SC-Ad is for administration by inhalation to infect a cell of the mammal thereby causing expression of the immunogen without production of new infectious virions, thus inducing the immune response

[0032] In a suitable embodiment, the SC-Ad is formulated as a liquid formulation. In a suitable embodiment, the liquid formulation comprises NaCl, histidine, MgC12, EDTA, polysorbate 80, sucrose, and ethanol.

[0033] In a suitable embodiment, the SC-Ad is formulated as a solid powder.

[0034] In a suitable embodiment, the SC-Ad is administered in an aerosol.

[0035] In a suitable embodiment, the SC-Ad is administered in a therapeutically effective amount of at least 1 x 1010SC-Ad viral particles.

[0036] In a suitable embodiment, the SC-Ad is administered in a therapeutically effective amount of between 1 x 1010and 3 x 1011SC-Ad viral particles.

[0037] In a suitable embodiment, the administration of the SC-Ad by inhalation is provided as a vaccine boostto a mammal that has received a previous vaccination to the pathogen before said subsequent administration of the SC-Ad by inhalation.

[0038] In a suitable embodiment, the administration of the SC-Ad by inhalation is provided as a vaccine boost to a mammal that has received a previous vaccination to the pathogen at least 90 days before said subsequent administration of the SC-Ad by inhalation.

[0039] In a suitable embodiment, the prior vaccination did not comprise administration of the SC-Ad.

[0040] In a suitable embodiment, the prior vaccine comprised administration of the SC-Ad.

[0041] In a suitable embodiment, the pathogen is a respiratory pathogen.

[0042] In a suitable embodiment, the immunogen of the respiratory pathogen is selected from the group consisting of: a SARS-CoV-2 immunogen, a SARS-CoV immunogen, a HCoV NL63 immunogen, a HKU1 immunogen, a MERS-CoV immunogen, an influenza immunogen, a respiratory syncytial virus (RSV) immunogen, a metapneumovirus (MPV) immunogen, a rhinovirus immunogen, a bocavirus immunogen, a parainfluenza virus (PIV) immunogen, a Streptococcus pneumoniae immunogen, a Bordetella pertussis immunogen, a Haemophilus influenzae immunogen, a Mycobacterium tuberculosis immunogen, and a Klebsiella pneumonia immunogen.

[0043] In a suitable embodiment, the pathogen is a coronavirus, and the immunogen is a coronavirus immunogen comprisinga coronavirus Spike polypeptide, or an immunogenic fragment thereof

[0044] In a suitable embodiment, a mucosal immune response is induced.

[0045] In a suitable embodiment, the mucosal immune response leadsto an increase in the amount of serum IgG antibodies that bind to said immunogen.

[0046] In a suitable embodiment, the mucosal immune response leadsto an increase in the amount of IgA antibodies that bind to said immunogen.

[0047] In a suitable embodiment, the expression of the immunogen in said cell leads to an increase in the amount of serum IgA antibodies that bind to said immunogen

[0048] In a suitable embodiment, the expression of the immunogen in said cell leads to an increase in the amount of secretory IgA antibodies that bind to said immunogen

[0049] In a suitable embodiment, the SC-Ad comprises a genomelacking at least a portion of a nucleic acid sequence that encodes an adenovirus polypeptide, and wherein the SC-Ad comprises said adenovirus polypeptide.

[0050] In a suitable embodiment, the adenovirus polypeptide is selected from the group consisting of a fiber polypeptide, a V polypeptide, a hexon polypeptide, a penton base polypeptide, and a pllla polypeptide.

[0051] A pharmaceutical composition comprising:

[0052] • a SC-Ad comprising a nucleic acid sequence encoding an immunogen of a pathogen; and

[0053] • excipients suitable for use in administration of said SC-Ad to a mammal by inhalation. In a suitable embodiment the pharmaceutical composition comprises a liquid formulation.

[0054] In a suitable embodiment the liquid formulation comprises NaCl, histidine, MgCl2, EDTA, polysorbate 80, sucrose, and ethanol as excipients.

[0055] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although methods and materials similar or equivalent to those described herein can be used to practice the invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

[0056] The details of one or more embodiments of the invention are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims.

[0057] BRIEF DESCRIPTION OF THE FIGURES

[0058] Fig 1 - Fig 1 is a is a line graph plotting the fold increase of anti-coronavirus spike protein IgG antibodies detected of each volunteer in the clinical trial on the indicated days following administration of a heterologous SC-Ad6-l boost by inhalation.

[0059] Fig 2 - Fig 2 is a is a line graph plotting the average fold increase of anti-coronavirus spike protein IgG antibodies detected in humans on the indicated days following administration of a heterologous SC-Ad6-l boost by inhalation.

[0060] Fig 3 - Fig 3 is a is a line graph plotting the fold increase of anti-coronavirus RBD IgG antibodies detected of each volunteer in the clinical trial on the indicated days following administration of a heterologous SC-Ad6-l boost by inhalation. Fig 4 - Fig 4 is a is a line graph plotting the average fold increase of anti-coronavirus RBD IgG antibodies detected in humans on the indicated days following administration of a heterologous SC-Ad6-l boost by inhalation.

[0061] Fig 5 - Fig 5 is a is a line graph plotting the fold increase of anti -coronavirus nucleocapsid IgG antibodies detected of each volunteer in the clinical trial on the indicated days following administration of a heterologous SC-Ad6-l boost by inhalation.

[0062] Fig 6 - Fig 6 is a is a line graph plotting the average fold increase of anti-coronavirus nucleocapsidlgGantibodies detected in humans on the indicated day sfollowing administration of a heterologous SC-Ad6-l boost by inhalation.

[0063] Fig 7 - Fig 7 is a line graph plotting the average fold increase of anti-coronavirus spike protein IgG antibodies in serum for three different Cohorts on the indicated days following administration of a heterologous SC-Ad6-l boost by inhalation.

[0064] Fig 8 - Fig 8 A is a line graph plotting the averagefold increase of anti-coronavirus spike protein IgA antibodies in serum for three different Cohorts on the indicated days following administration of a heterologous SC-Ad6-l boost by inhalation; Fig 8B is a line graph plotting the average fold increase of anti-coronavirus spike protein IgAantibodies in samplesfrom nasal mucosa for three different Cohorts on the indicated days following administration of a heterologous SC-Ad6-l boost by inhalation; and Fig 8C is a line graph plotting the average fold increase of anti-coronavirus spike protein IgA antibodies in saliva for three different Cohorts on the indicated days following administration of a heterologous SC-Ad6-l boost by inhalation.

[0065] DETAILED DESCRIPTION OF THE INVENTION

[0066] The inventors have identified new and advantageous methods of treatment and medical uses in which SC-Ads are delivered by inhalation to immunize mammals. The SC-Ads are used to deliver one or more immunogens (e.g., one or more immunogens associated with a pathogen causing an infection) to cells within a mammal (such as a human). As a result, the mammal produces an effective immune response against the immunogen(s), and hence the pathogen. In particular, the methods of treatment and medical uses of the invention facilitate production of therapeutically effective IgG and / or IgA and / or T cell immune responses against the immunogen(s) and so against the pathogen.

[0067] As set out in the Examples, the methods of treatment and medical uses disclosed herein are of particular utility in clinical situations in which it is desirable to induce a systemic immune response. The methods of treatment and medical uses of the invention are well suited to use to generate high levels of antibodies in the serum of a recipient mammal. The methods of treatment and medical uses of the invention are able to generate high levels of IgG antibodies in the serum of a recipient mammal and / or high levels of IgA antibodies in the serum of a recipient mammal. The inventors having newly identified the ability of the methods of treatment and medical uses of the invention to generate immune responses of this sort, the skilled person will readily be able to identify clinical contexts in which such responses may be desirable.

[0068] The immune response generated may be a mucosal immune response. In particular, the immune response may be a mucosal response of the respiratory mucosa. Such a response is particularly effective in providing protection from respiratory pathogens.

[0069] Single cycle adenovirus vectors for use in method of treatment or medical uses of the invention

[0070] The first aspect of the invention provides a method for inducing an immune response against a pathogen in a mammal, the method comprising administering a single cycle adenovirus (SC- Ad) by inhalation to said mammal, wherein said SC-Ad comprises a nucleic acid sequence encoding an immunogen of said pathogen, and infects a cell of said mammal thereby causing expression of said immunogen leading to the induction of said immune response without production of new infectious virions.

[0071] The second aspect of the invention provides a single cycle adenovirus (SC-Ad) comprising a nucleic acid sequence encoding an immunogen of to a pathogen for use in inducing an immune response against the pathogen in a mammal, wherein the SC-Ad is administered by inhalation and infects a cell of the mammal thereby causing expression of the immunogen leading to the induction of the immune response without production of new infectious virions. A SC-Ad suitable for use in the methods of treatment or medical uses of the invention differs substantially from a typical adenovirus vector. In particular, the SC-Ads for use in the methods of treatment or medical uses of the invention are engineered so that they are only able to undergo a single round of replication within host cells. This is in contrast to traditional adenovirus vectors, in which the virus is capable of multiple rounds of replication, or “replication defective” adenovirus vectors (also known as “replication deficient” vectors) which do not replicate at all in the host.

[0072] The continuing replication of traditional adenovirus vectors plays an important role in their use as vaccines, allowing them to generate large quantities of immunogens that give rise to the protective immune response. However, such ongoing replication can potentially cause safety concerns. To address this, SC-Ads have been developed that incorporate genomic changes preventing them from producing infectious virions.

[0073] Adenovirus vectors typically encode several genes that are essential for viral replication, including those involved in DNA replication, capsid formation, and virion assembly. In a suitable embodiment of an SC-Ad suitable for use in the methods of treatment or medical uses of the invention, one or more of these essential genes are deleted or inactivated. Without these genes, the vector is unable to complete a full replication cycle and produce infectious virions. However, if the corresponding adenovirus polypeptide is provided within the adenovirus, this allows the virus to overcome the lack of the deleted or inactivated gene. The adenovirus vectors are thereby enabled to undergo a single cycle of expression of the immunogen, without production of infectious virions.

[0074] Thus, SC-Ads suitable for use in the methods of treatment or medical uses of the invention have several advantages over other typical viral vectors e.g., other adenovirus vectors. Since SC-Ads suitable for use in the methods of treatment or medical uses of the invention are only able to undergo a single round of replication, and not multiple rounds of replication within host cells, this reduces or removes the risk of uncontrolled production of infectious virions and limits potential adverse effects associated with viral replication, such as cytotoxicity and immunogenicity. Consequently, they are generally considered safer for therapeutic use.

[0075] Routes of administration When practicing the methods of treatment or medical uses of the invention, SC-Ads are provided to a subjectby inhalation. Suitably a SC-Adis administered to a subjectby inhalation through the mouth (suitably as part of a pharmaceutical composition of the invention), f ollowed by exhalation of breath containing the SC-Ads via the mouth and / or nose.

[0076] Inhalation as a route of administration allows mucosal delivery of the SC-Ads. In particular, inhalation enables provision of a SC-Ad suitable for use in the methods of treatment or medical uses of the invention to the respiratory mucosa (e.g., epithelial surfaces) of the respiratory system.

[0077] The SC-Ad may particularly be provided to the lower respiratory system. The lower respiratory system as used herein refers to the trachea, lungs and the oropharynx. More specifically, the lower respiratory system includes the trachea, bronchi, bronchioles, alveoli, lungs and the oropharynx. Thus, administration of a SC-Ad suitable for use in the methods of treatment or medical uses of the invention by inhalation facilitates delivery of a SC-Ad directly to the respiratory mucosa to induce an immune response.

[0078] The respiratory mucosa, also known as the respiratory epithelium or respiratory lining, is a specialized mucous membrane that lines components of the respiratory system. It consists of a layer of epithelial cells supported by connective tissue and contains various immune cells that play important roles in respiratory defence and immune surveillance. It will be appreciated that respiratory pathogens frequently enter a subject by means of the respiratory mucosa, and so the ability to provide SC-Ad to this tissue, and thereby generate a strong mucosal immune response within the respiratory mucosa, is particularly advantageous.

[0079] Thus, administration of a SC-Ad suitable for use in the methods of treatment or medical uses of the invention by inhalation has multiple advantages over other forms of administration that are particularly relevant to, but not limited to, use in the generation of an immune response against a respiratory pathogen.

[0080] For example, administration by inhalation can provide targeted delivery to the respiratory system. By delivering a SC-Ad suitable for use in the methods of treatment or medical uses of the invention directly to the respiratory system, it can induce a robust immune response at a site of infection, or of potential infection, thus providing better protection against respiratory diseases.

[0081] Additionally, administration by inhalation can induce significant local immune responses. The respiratory mucosa is rich in immune cells and mucosal-associated lymphoid tissue (MALT), which play a crucial role in mucosal immunity. Administration by inhalation can stimulate local immune responses to the pathogen in the respiratory mucosa, including the production of secretory IgA antibodies and the activation of mucosal immune cells, such as T cells and dendritic cells.

[0082] Furthermore, administration by inhalation is considered more user friendly. Administration by inhalation offer a needle-free alternative to traditional injectable vaccines. This can improve acceptance and compliance, especially in paediatric populations.

[0083] Inhalation constitutes a favourable route for administering agents, such as an SC-Ad suitable for use in the methods of treatment or medical uses of the invention, targeting respiratory pathogens. Inhalation allows agents to be directly delivered to the site of infection in the respiratory tract, such as the lungs and airways, where respiratory pathogens primarily reside and cause infections. This targeted delivery enables higher agent concentrations atthe infection site, increasing the efficacy of treatment while minimizing systemic side effects. Inhalation also provides rapid delivery of agents to the lungs, allowing for quick absorption and onset of action. This is particularly beneficial in treating acute respiratory infections caused by pathogens such as viruses or bacteria, where rapid symptom relief and control of infection are crucial. Additionally, inhalation administration minimizes systemic exposure to the agent compared to oral or intravenous routes. This reduces the risk of systemic side effects and toxicity while maximizing the therapeutic effect at the site of infection in the respiratory tract.

[0084] Inhalation administration often requires lower drug doses compared to systemic routes, as drugs delivered directly to the lungs can achieve higher local concentrations. This can lead to improved patient compliance and reduced drug costs.

[0085] Inhalation administration is distinguished from other mucosal administration routes, such as intranasal and oral administration, based on the specific anatomical and physiological characteristics of the respiratory tract and the delivery method used. For example, for inhalation the SC-Ad is delivered directly to the respiratory tract, typically to the lungs, via inhalation of aerosolized particles or droplets whereas for intranasal or oral administration the SC-Ad would be administered primarily via nasal cavity, through the mouth and swallowed, entering the gastrointestinal tract.

[0086] Furthermore, in administration via inhalation the primary target tissues are the respiratory epithelium and alveoli within the lungs, in contrast, intranasal or oral administration primarily targets the nasal mucosa and adjacenttissues or the gastrointestinal tract, including the stomach and intestines.

[0087] Additionally, administration via inhalation means that absorption of the drug occurs primarily via diffusion across the respiratory epithelium and alveolar membranes into the bloodstream. Intranasal or oral administration typically facilitate absorption via diffusion across the nasal mucosa into the bloodstream or via drainage into the nasopharynx and subsequent swallowing into the gastrointestinal tract or straight to the gastrointestinal tract.

[0088] Administration of an SC-Ad suitable for use in the methods of treatment or medical uses of the invention by inhalation comprises delivery of the SC-Ad to the respiratory mucosa of the respiratory system. Preferably, administration of an SC-Ad suitable for use in the methods of treatment or medical uses of the invention by inhalation comprises delivery to the respiratory mucosa of the lower respiratory system.

[0089] In a suitable embodiment, administration of a SC-Ad suitable for use in the methods of treatment or medical uses of the invention by inhalation comprises delivery of said SC-Ad primarily to the respiratory mucosa of the lower respiratory system. In a suitable embodiment administration of a SC-Ad suitable for use in the methods of treatment or medical uses of the invention by inhalation comprises delivery of said SC-Ad to the respiratory mucosa of the lower respiratory system.

[0090] In a suitable embodiment, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention is administered in an aerosol. An aerosol refers to a suspension of fine solid or liquid particles in a gas. In a suitable embodiment, administration of a SC-Ad suitable for use in the methods of treatment or medical uses of the invention by inhalation is via an inhalation device. Any suitable inhalation device may be used and a person skilled in the art would be aware of such devices including inhalers and nebulisers.

[0091] In a suitable embodiment, the inhalation device is a nebuliser or inhaler. Any suitable nebuliser or inhaler may be used for the administration of a SC-Ad suitable for use in the methods of treatment or medical uses of the invention and a person skilled in the art would be aware of such nebulisers or inhalers.

[0092] In a suitable embodiment, the inhaler is selected from the group consisting of: Metered-Dose Inhaler (MDI), Dry Powder Inhaler (DPI) and Soft Mist Inhaler (SMI). Suitably, the inhaler is a metered-dose inhaler. Suitably, the inhaler is a dry powder inhaler. Suitably, the inhaler is a soft mist inhaler.

[0093] In a suitable embodiment, the nebuliser is selected from the group consisting of: mesh nebuliser, ultrasonic nebuliser, jet nebuliser and breath-actuated nebuliser. Suitably, the nebuliser is a mesh nebuliser. Suitably, the nebuliser is an ultrasonic nebuliser. Suitably, the nebuliser is a jet nebuliser. Suitably, the nebuliser is a breath-actuated nebuliser.

[0094] In some cases, the route and / or mode of administration of a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can be adjusted for the mammal being treated.

[0095] Method of treatment and medical uses

[0096] The first aspect of the invention provides a method for inducing an immune response against a pathogen in a mammal, the method comprising administering a single cycle adenovirus (SC- Ad) by inhalation to said mammal, wherein said SC-Ad comprises a nucleic acid sequence encoding an immunogen of said pathogen, and infects a cell of said mammal thereby causing expression of said immunogen leading to the induction of said immune response without production of new infectious virions.

[0097] The second aspect of the invention provides a single cycle adenovirus (SC-Ad) comprising a nucleic acid sequence encoding an immunogen of a pathogen for use in inducing an immune response against the pathogen in a mammal, wherein the SC-Ad infects a cell of the mammal thereby causing expression of the immunogen leading to the induction of the immune response without production of new infectious virions.

[0098] Inducing an immune response

[0099] For the purposes of the present disclosure, induction of an immune response refers to the activation and stimulation of the body's immune system to recognize and mount a defence against a pathogen (e.g., viruses, bacteria, fungi) in response to provision of an immunogen that is associated with the pathogen and is encoded by a nucleic acid in an SC-Ad.

[0100] In particular, inducing an immune response in the context of the present invention involves triggering one or more components of the immune system selected from the group consisting of: immune cells such as T cells, B cells, and antigen-presenting cells, as well as signalling molecules such as cytokines and antibodies. Suitably, the immune response induced by the methods of treatment or medical uses of the invention may be a pro-inflammatory immune response, as considered further below.

[0101] This document provides methods of treatment and medical uses using SC-Ads to induce an immune response against a pathogen. SC-Ads suitable for use in the methods of treatment or medical uses of the invention, methods of treatment or medical uses of the invention can increase an immune response (e.g., an increased antibody response, such as an increased IgA response) against a pathogen (e.g., a bacterial or a viral pathogen associated with an immunogen encoded by the SC-Ad).

[0102] For example, SC-Ads suitable for use in the methods of treatment or medical uses of the invention, methods of treatment or medical uses of the invention may be used to induce in a mammal an immune response effective to reduce the severity of an infection caused by a pathogen associated with the immunogen(s) encoded by the SC-Ads.

[0103] In some cases, SC-Ads suitable for use in the methods of treatment or medical uses of the invention, methods of treatment or medical uses of the invention may be used to induce in the mammal an immune response effective to prevent the mammal from exhibiting symptoms of an infection caused by a pathogen associated with the immunogen(s) encoded by the SC-Ads. In some embodiments, SC-Ads suitable for use in the methods of treatment or medical uses of the invention, when used in the methods of treatment or medical uses of the invention, may induce a mucosal immune response. A mucosal immune response refers to the specialized branch of the immune system that provides defence against pathogens and maintains immune homeostasis at mucosal surfaces throughout the body. Mucosal surfaces are found in various organs, including the respiratory tract, gastrointestinal tract, genitourinary tract, and ocular surfaces, and they are constantly exposed to a wide range of environmental challenges, including pathogens, toxins, and commensal microorganisms. The methods of treatment or medical uses of the invention may induce a mucosal immune response in the mucosa of the respiratory tract.

[0104] The mucosal immune response involves a complex interplay of various immune cells and molecules to protect mucosal surfaces. The methods of treatment or medical uses of the invention may induce a mucosal immune response to the pathogen involving one or more of B-cells, T-cells, IgA antibodies and IgG antibodies.

[0105] In some cases, SC-Ads suitable for use in the methods of treatment or medical uses of the invention, methods of treatment or medical uses of the invention may increase a B cell response to a pathogen within a mammal. For example, SC-Ads suitable for use in the methods of treatment or medical uses of the invention, methods of treatment or medical uses of the invention may increase the number of activated B cells (e.g., plasmablasts, plasma cells, and memory B cells) within the mammal by at least 10%, at least 11 %, at least 12%, at least 13 %, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21%, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least28%, at least29%, at least 30%, at least 31%, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least 40%, at least 41 %, at least 42%, at least 43%, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least49%, at least 50%, at least 51%, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61 %, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even by 100%. Indeed, a suitable increase may be by at least twofold, at least 5-fold, at least 10-fold, at least 25-fold, at least 50-fold, or at least 100-fold or more.

[0106] In some cases, SC-Ads suitable for use in the methods of treatment or medical uses of the invention, methods of treatment or medical uses of the invention may increase a number of antibodies (e.g., antibodies against a pathogen such as abacterial or a viral pathogen associated with the immunogen(s) encoded by the SC-Ads) within a mammal. For example, SC-Ads suitable for use in the methods of treatment or medical uses of the invention, methods of treatment or medical uses of the invention may increase the number of antib odies to a pathogen within a mammal by at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21%, at least

[0107] 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least

[0108] 29%, at least 30%, at least 31%, at least 32%, at least 33%, at least 34%, at least 35%, at least

[0109] 36%, at least 37%, at least 38%, at least 39%, at least 40%, at least 41 %, at least 42%, at least

[0110] 43%, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least

[0111] 50%, at least 51%, at least 52%, at least 53%, atleast 54%, atleast 55%, atleast 56%, at least 57%, at least 58%, at least 59%, at least 60%, atleast 61%, atleast 62%, atleast 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least

[0112] 71%, at least 72%, at least 73%, at least 74%, atleast 75%, atleast 76%, atleast 77%, at least

[0113] 78%, at least 79%, at least 80%, at least 81%, atleast 82%, atleast 83%, atleast 84%, at least 85%, at least 86%, at least 87%, at least 88%, atleast 89%, atleast 90%, atleast 91%, at least

[0114] 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least

[0115] 99%, or even by 100%. Indeed, a suitable increase may be by at least two-fold, atleast 5 -fold, at least 10-fold, at least 25-fold, atleast 50-fold, or at least 100-fold or more. Suitably, such antibodies may be antibodies of a neutralising immune response. The nature of antibodies as constituting part of a neutralising immune response may be determined by suitable assays in vitro.

[0116] In some cases, SC-Ads suitable for use in the methods of treatment or medical uses of the invention, methods of treatment or medical uses of the invention may be used to produce antibodies against a immunogen(s) within a mammal for, for example, about one week (e.g., from about 1 to about 7 days, from about 1 to about 6 days, from about 1 to about 5 days, from about 1 to about 4 days, from about 1 to about 3 days, from about 2 to about 7 days, from about 3 to about 7 days, from about 4 to about 7 days, from about 5 to about 7 days, from about 2 to about 6 days, from about 3 to about 5 days, from about 2 to about 4 days, from about 3 to about

[0117] 5 days, or from about 4 to about 6 days).

[0118] In a suitable embodiment, SC-Ads suitable for use in the methods of treatment or medical uses of the invention, methods of treatment or medical uses of the invention may increase the amount of serum IgG antibodies thatbind to an immunogen encoded by the SC-Ads within a mammal. For example, SC-Ads suitable for use in the methods of treatment or medical uses of the invention, methods of treatment or medical uses of the invention may increase the amount of serum IgG antibodies thatbind to an immunogen encoded by the SC-Ads within a mammal by at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21 %, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31%, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least 40%, at least 41 %, at least 42%, at least 43 %, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51%, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even by 100%. Indeed, a suitable increase may be by at least two-fold, at least 5-fold, at least 10-fold, at least 25-fold, at least 50-fold, or at least 100-fold or more.

[0119] In a suitable embodiment, SC-Ads suitable for use in the methods of treatment or medical uses of the invention, methods of treatment or medical uses of the invention may increase the amount of IgAantibodies thatbind to an immunogen encoded by the SC-Ads within a mammal. For example, SC-Ads suitable for use in the methods of treatment or medical uses of the invention, methods of treatment or medical uses of the invention may increase the amount of IgA antibodies thatbind to an immunogen encoded by the SC-Ads within a mammal by at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21%, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31%, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least 40%, at least 41%, at least 42%, at least 43%, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51%, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even by 100%. Indeed, a suitable increase may be by at least two-fold, at least 5-fold, at least 10-fold, at least 25 -fold, at least 50-fold, or at least 100-fold or more.

[0120] In a suitable embodiment, SC-Ads suitable for use in the methods of treatment or medical uses of the invention, methods of treatment or medical uses of the invention may increase the amount of serum IgA antibodies that bind to an immunogen encoded by the SC-Ads within a mammal. For example, SC-Ads suitable for use in the methods of treatment or medical uses of the invention, methods of treatment or medical uses of the invention may increase the amount of serum IgA antibodies that bind to an immunogen encoded by the SC-Ads within a mammal by at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21 %, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31%, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least 40%, at least 41 %, at least 42%, at least 43 %, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51%, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even by 100%. Indeed, a suitable increase may be by atleasttwo-fold, at least 5 -fold, at least 10-fold, at least 25-fold, at least 50-fold, or at least 100-fold or more.

[0121] In a suitable embodiment, SC-Ads suitable for use in the methods of treatment or medical uses of the invention, methods of treatment or medical uses of the invention may increase the amount of secretory IgA antibodies that bind to an immunogen encoded by the SC-Ads within a mammal. For example, SC-Ads suitable for use in the methods of treatment or medical uses of the invention, methods of treatment or medical uses of the invention may increase the amount of secretory IgA antibodies that bind to an immunogen encoded by the SC-Ads within a mammal by atleast 10%, at least 11%, atleast 12%, at least 13%, atleast 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21 %, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31%, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least 40%, at least 41 %, at least 42%, at least 43 %, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51%, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even by 100%. Indeed, a suitable increase may be by atleasttwo-fold, atleast 5 -fold, atleast 10-fold, at least 25-fold, at least 50-fold, or at least 100-fold or more.

[0122] In some cases, SC-Ads suitable for use in the methods of treatment or medical uses of the invention, methods of treatment or medical uses of the invention may increase a T cell response to the pathogen within a mammal. For example, SC-Ads suitable for use in the methods of treatment or medical uses of the invention, methods of treatment or medical uses of the invention may increase the number of activated T cells (e.g., cytotoxic T cells and macrophages) within the mammal by at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least21%, at least22%, at least 23%, at least 24%, at least25%, at least26%, at least27%, at least28%, at least29%, at least 30%, at least 31%, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least40%, at least41%, at least 42%, at least 43%, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least49%, at least 50%, at least 51%, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even by 100%. Indeed, a suitable increase may be by at least twofold, at least 5-fold, at least 10-fold, at least 25-fold, at least 50-fold, or at least 100-fold or more.

[0123] The immune response induced by the methods of treatment or medical uses of the invention may be a pro-inflammatory immune response. Characteristics of pro-inflammatory immune responses will be well known to those of skill in the art. Without limitation, they may include expression of one, more or all of the following molecules: TNF-a, IFN-y, IL-1, IL-6, IL-12, IL-18, and GM-CSF. For example, SC-Ads suitable for use in the methods of treatment or medical uses of the invention, methods of treatment or medical uses of the invention may increase a pro-inflammatory immune response within the mammal by at least 10%, at least 11%, atleast 12%, at least 13%, at least 14%, at least 15%, at least 16%, atleast 17%, at least

[0124] 18%, at least 19%, at least 20%, at least 21 %, at least 22%, at least 23%, at least 24%, at least

[0125] 25%, at least26%, at least 27%, at least 28%, atleast 29%, atleast 30%, atleast 31%, at least

[0126] 32%, at least 33%, at least 34%, at least 35%, atleast 36%, atleast 37%, atleast 38%, at least

[0127] 39%, at least40%, at least 41%, at least 42%, atleast 43%, atleast 44%, atleast 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51%, at least 52%, at least

[0128] 53%, at least 54%, at least 55%, at least 56%, atleast 57%, atleast 58%, atleast 59%, at least

[0129] 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least

[0130] 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least

[0131] 74%, at least 75%, at least 76%, at least 77%, atleast 78%, atleast 79%, atleast 80%, at least

[0132] 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least

[0133] 88%, at least 89%, at least 90%, at least 91%, atleast 92%, atleast 93%, atleast 94%, at least

[0134] 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even by 100%. Indeed, a suitable increase may be by at least two-fold, at least 5-fold, at least 10-fold, at least 25-fold, at least 50-fold, or at least 100-fold or more. Expression of markers characteristic of a pro- inflammatory immune response may be investigated and quantified by any suitable means, such as ELISPOT.

[0135] The immune response induced by the methods of treatment or medical uses of the invention may increase migration of immune cells, such as T cells, into the lungs, as part of a protective immune response For example, SC-Ads suitable foruse in the methods oftreatment or medical uses of the invention, methods of treatment or medical uses of the invention may increase migration of immune cells, such as T cells, into the lungs of the mammal by at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21%, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31 %, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least40%, at least 41%, at least 42%, at least43%, at least 44%, at least45%, at least46%, at least47%, at least 48%, at least 49%, at least 50%, at least 51%, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even by 100%. Indeed, a suitable increase may be by at least two-fold, at least 5-fold, at least 10-fold, at least 25-fold, at least 50-fold, or at least 100-fold or more. Migration of immune cells, such asT cells, into the lungs may be investigated and quantified by any suitable means, such as bronchoalveolar lavage (BAL).

[0136] As illustrated in the Examples, the methods of treatment and medical uses of the invention may be of use in clinical situations in which it is desirable to increase the levels of antibodies (such as serum antibodies) to the encoded immunogen by atleast0.5-fold, at least 1 -fold, at least 1.5- fold, at least 2-fold, at least 2.5 -fold, at least 3 -fold, at least 3.5 -fold, atleast 4-fold, atleast4.5- fold, at least 5 -fold, at least 5.5 -fold, at least 6-fold, at least 6.5 -fold, at least 7 -fold, at least 7.5- fold, at least 8-fold, at least 8.5-fold, atleast 9-fold, at least 9.5-fold, atleast 10-fold, at least 10.5-fold, or more, as compared to the baseline levels of such antibodies before administration of SC-Ads by inhalation.

[0137] In the case that the methods of treatment or medical uses of the invention are being used to provide a first incidence of vaccination (whether as a prime before a subsequent boost, or as the sole incidence of vaccination) the baseline may be indicative of an unvaccinated level of such antibodies. In the case that the methods of treatment or medical uses of the invention are being used to provide a second or further incidence ofvaccination (for example as a vaccination boost, whether a heterologous boost or homologous boost) the baseline may be indicative of a vaccinated level of such antibodies prior to boost.

[0138] In keeping with this, the inventors have demonstrated that methods of treatment and medical uses in accordance with the invention may be of use in clinical situations in which it is desired to induce a systemic immune response characterised by the presence of serum antibodies to the encoded immunogen. Methods of treatment and medical uses in accordance with the invention may be of use in clinical situations in which it is desired to induce a systemic immune response characterised by the presence of high levels and / or elevated levels of serum antibodies to the encoded immunogen (in keeping with the increased fold changes of antibodies discussed above).

[0139] For example, the inventors have demonstrated that methods of treatment and medical uses in accordance with the invention may be of use in clinical situations in which it is desired to preferentially induce a systemic immune response characterised by the presence of serum antibodies to the encoded immunogen. Suitably, such a response may be characterised by the the presence of a level of serum antibodies (e.g. IgG and / or IgA antibodies) that is increased with respect to the baseline in line with the suggestions set out above.

[0140] The inventors have demonstrated that methods of treatment and medical uses in accordance with the invention may be of use in clinical situations in which it is desired to induce a prolonged systemic immune response characterised by the presence of serum antibodies to the encoded immunogen.

[0141] As set out further in the Examples, the inventors have demonstrated that methods of treatment and medical uses in accordance with the invention may be of use in clinical situations in which it is desired to preferentially induce a systemic immune response characterised by the presence of increased levels of IgG and / or IgA serum antibodies to the encoded immunogen. Such increases may, for example, be determined with respect to the fold changes outlined above.

[0142] Subjects or patients

[0143] The methods of treatment of the invention are practiced in respect of mammalian subjects or patients requiring the induction of an immune response against a pathogen. The words “subject” and “patient” may be used interchangeably in the context of the present disclosure.

[0144] Suitably, a subject or patient requires induction of an immune response against a respiratory pathogen.

[0145] The subject may have symptoms of a disease or condition caused by a pathogen requiring induction of an immune response for treatment. In such cases the subject may have an acute disease, or a chronic disease. Alternatively, the subject may be identified as being at elevated risk of developing a disease requiring induction of the immune response for prevention.

[0146] Mammals

[0147] The methods of treatment or medical uses of the invention make use of SC-Ads, as defined herein, to induce an immune response in a mammal. The SC-Ads can be administered to any appropriate mammal (e.g., to increase an immune response against a pathogen such as a bacterial or a viral pathogen associated with the immunogen(s) encoded by the SC-Ads within that mammal).

[0148] In some cases, the mammal can be a mammal that has not had a previous infection with a pathogen associated with an immunogen encoded by a SC-Ad suitable for use in the methods of treatment or medical uses of the invention. In some cases, the mammal can be a mammal that has had a previous infection with a pathogen closely related (e.g., genetically related) to a pathogen associated with an immunogen encoded by an adenovirus vector provided herein.

[0149] In some cases, the mammal can be a mammal that has had a previous infection with a pathogen associated with an immunogen encodedby a SC-Ad suitable for use in the methods of treatment or medical uses of the invention. In some cases, the mammal can be a mammal that has an infection (e.g., an ongoing infection) with a pathogen associated with an immunogen encoded by a SC-Ad suitable for use in the methods of treatment or medical uses of the invention.

[0150] Examples of mammals that can be administered SC-Ads suitable for use in the methods of treatment or medical uses of the invention include, without limitation, humans, non-human primates such as monkeys or chimpanzees, domestic animals (such as dogs, cats, mice, rats, rabbits, and hamsters), bats, raccoons, ferrets, and agricultural animals (such as horses, cows, pigs, and sheep).

[0151] In some cases, a mammal (such as a human) can be administered one or more SC-Ads suitable for use in the methods of treatment or medical uses of the invention to increase an immune response against a pathogen (e.g., a bacterial or a viral pathogen associated with an immunogen encoded by the SC-Ad).

[0152] Treatment Regimes

[0153] SC-Ads suitable for use in the methods of treatment or medical uses of the invention can be administered to a mammal (e.g., a human) in any appropriate frequency. The frequency of administration can be any frequency that can induce an immune response in a mammal without producing significant toxicity to the mammal.

[0154] In some cases, SC-Ads suitable for use in the methods of treatment or medical uses of the invention can be administered to a mammal once (e.g., in a single administration) in the methods of treatment or medical uses of the invention.

[0155] In some cases, SC-Ads suitable for use in the methods of treatment or medical uses of the invention can be administered to a mammal several times (e.g., as several administrations) in the methods of treatment or medical uses of the invention. For example, the frequency of administration can be from about once a day to about every three days, from about once a day to about once a week, from about once a week to about every 3 weeks, or from about once a week to about every 6 weeks. The frequency of administration can remain constant or can be variable during the duration of treatment. As with the effective amount discussed below, various factors can influence the actual frequency of administration used for a particular application. For example, the effective amount, duration of treatment, useof multiple treatment agents, and / or route of administration may require an increase or decrease in administration frequency.

[0156] SC-Ads suitable for use in the methods of treatment or medical uses of the invention can be administered to a mammal (e.g., a human) for any appropriate duration in the methods of treatment or medical uses of the invention. An effective duration for administering or using a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can be any duration that can induce an immune response in a mammal without producing significant toxicity to the mammal. For example, the effective duration can vary from a couple of days to one week, from several days to several weeks, or from a few days to a month. Multiple factors can influence the actual effective duration used for a particular treatment. For example, an effective duration can vary with the frequency of administration, effective amount, use of multiple treatment agents, and / or route of administration.

[0157] In some cases, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can be administered to a mammal (e.g., a human) at an effective amount one, two, or three times with one week to four weeks between each administration when more than one administration is used.

[0158] In some cases, one or more SC-Ads suitable for use in the methods of treatment or medical uses of the invention can be administered to a mammal (e.g., a human) in the methods of treatment or medical uses of the invention as the sole active ingredient to induce an immune response (e.g., an increased antibody response) against a pathogen (e.g., a bacterial or a viral pathogen associated with an immunogen encodedby the SC-Ads). For example, a composition containing SC-Ads suitable for use in the methods of treatment or medical uses of the invention can be administered to a mammal (e.g., a human) as the sole active ingredient to induce an immune response (e.g., an increased antibody response) against a coronavirus.

[0159] In some cases, one or more SC-Ads suitable for use in the methods of treatment or medical uses of the invention can be administered to a mammal (e.g., a human) in the methods of treatmentor medical uses ofthe invention together with one or more (e.g., one, two, three, four, five or more) additional agents / therapiesusedto induce an immuneresponse (e.g., an increased antibody response) against a pathogen (e.g., a bacterial or a viral pathogen associated with an immunogen encoded by the SC-Ads). For example, a composition containing SC-Ads suitable for use in the methods of treatment or medical uses of the invention can be administered to a mammal (e.g., a human) together with one or more (e.g., one, two, three, four, five or more) additional agents / therapies used to induce an immune response (e.g., an increased antibody response) against a coronavirus. In cases where one or more SC-Ads suitable for use in the methods of treatment or medical uses of the invention are used in combination with one or more additional agents / therapies used to induce an immune response (e.g., an increased antibody response), the one or more SC-Ads suitable for use in the methods of treatment or medical uses of the invention and the one or more additional agents / therapies can be administered atthe same time (e.g., in a single composition) or independently. For example, one or more SC-Ads suitable for use in the methods of treatment or medical uses of the invention adenovirus can be administered first, and the one or more additional agents / therapies administered second, or vice versa.

[0160] In a suitable embodiment, the administration of a SC-Ad suitable for use in the methods of treatment or medical uses of the invention is provided as a vaccine boost to a mammal that has received a previous vaccination to the pathogen before the subsequent administration of the SC-Ad by inhalation. A vaccination boost, also known as a booster dose or booster shot, refers to an additional dose of a vaccine administered after the initial vaccination series. The purpose of a booster dose is to enhance and prolong the protective immune response induced by the primary vaccination series.

[0161] In a suitable embodiment of a “prime boost” vaccination treatment regimen, the further vaccination “boost” may be administered at least one week, at least two weeks, at least three weeks, at least four weeks, at least five weeks, or at least six weeks after the initial vaccination “prime”. For example, the further vaccination boost may be administered at least 50 days, at least 60 days, at least 70 days, at least 80 days, at least 90 days, at least 100 days, at least 110 days, at least 120 days, at least 130 days, at least 140 days, or at least 150 days after the initial vaccination prime.

[0162] Thus, the previously received vaccine can be one thatthe mammal received from about 14 days to about 120 months (e.g., e.g., from about 21 days to about 60 months, from about 21 days to about 36 months, from about 21 days to about 24 months, from about 21 days to about 12 months, from about 21 days to about 8 months, from about 3021 days to about 6 months, from about 21 days to about 4 months, from about 21 days to about 3 months, from about 21 days to about 2 months, from about 1 month to about 120 months, from about 3 months to about 120 months, from about 6 months to about 120 months, from about 12 months to about 120 months, from about 18 months to about 120 months, from about 24 months to about 120 months, from about 36 months to about 120 months, from about 48 months to about 120 months, from about 1 month to about 60 months, from about 2 months to about 48 months, from about 3 months to about 36 months, from about4 monthsto about 24 months, from about 6 months to about 18 months, from about 1 month to about 3 months, from about 3 months to about 6 months, from about 6 months to about 12 months, from about 12 months to about24 months, from about 24 months to about 48 months, from about 36 months to about 72 months, from about48 monthsto about 84 months, orfrom about60 monthsto about96 months)before the mammal is administered a SC-Ad suitable for use in the methods of treatment or medical uses of the invention (e.g., a SC-Ad engineered to encode a SARS-CoV-2 Spike polypeptide). A method of treatment or medical use of the invention may b e used as a booster at least 90 days after a previous vaccination for the pathogen.

[0163] In a suitable embodiment, the prior vaccination did not comprise administration of an SC-Ad suitable for use in the methods of treatment or medical uses of the invention. This approach is referred to as heterologous or mixed-dose vaccination boost. For example, SC-Ads suitable for use in the methods of treatment or medical uses of the invention which were designed to express a SARSCoV-2 polypeptide such as a spike polypeptide (or a fragment thereof) may be used as a heterologous boost to a human who received a prior mRNA-based or DNA-based SARS- CoV-2 vaccine (e.g., a Pfizer® vaccine such as Comirnaty® or an AstraZeneca® vaccine such as Vaxzevria®). In some cases, SC-Ads suitable for use in the methods of treatment or medical uses of the invention which are administered by inhalation can be used as a heterologous vaccination boost to deliver nucleic acid encoding one or more (e.g., one, two, three, four, five, six, seven, eight, nine, ten, or more) immunogens in a manner that triggers IgG, IgA, and T cell immune responses within a mammal (e.g., a human) that received at least one prior vaccine (e.g., one or more prior mRNA vaccines) against the targeted pathogen.

[0164] In a suitable embodiment, the prior vaccine comprised administration of an SC-Ad in the methods of treatment or medical uses of the invention. This approach is referred to as homologous vaccination boost. For example, SC-Ads suitable for use in the methods of treatment or medical uses of the invention may be used as a vaccine boost to a mammal that has received a previous vaccination to the pathogen at least 90 days before the subsequent administration of the SC-Ad by inhalation, wherein the previous vaccine was an SC-Ad based vaccination. The previously received SARS-CoV-2 vaccine can be one that the mammal received at least 90 days (e.g., at least 120 days, at least 150 days before said administering, or at least 180 days) before the mammal is administered a SC-Ad suitable for use in the methods of treatment or medical uses of the invention (e.g., a SC-Ad engineered to encode a SARS- CoV-2 Spike polypeptide).

[0165] In some cases, SC-Ads suitable for use in the methods of treatment or medical uses of the invention can be administered by inhalation to a mammal (e.g., a human) as a heterologous or homologous vaccination boost to increase IgG, IgA, and T cell immune responses within the mammal. In some cases, SC-Ad suitable for use in the methods of treatment or medical uses of the invention can be administered as a heterologous or homologous vaccination boost to a mammal (e.g., a human) that previously received a vaccine targeting the same pathogen (e.g., a bacterial or a viral pathogen) to increase the number of IgG and IgA antibodies (e.g., IgG and IgA antibodies against the pathogen associated with the immunogen(s) encoded by the SC- Ads) within the mammal.

[0166] For example, SC-Ads suitable for use in the methods of treatment or medical uses of the invention canbe administered by inhalation as a heterologous or homologous vaccination boost to a mammal (e.g., a human) that previously received a vaccine targeting the same pathogen (e.g., a bacterial or a viral pathogen) to increase the number of IgG and IgA antibodies within the mammal by at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21%, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31%, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least 40%, at least 41 %, at least 42%, at least 43%, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51%, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, atleast 61%, atleast 62%, atleast 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, atleast 75%, atleast 76%, atleast 77%, at least 78%, at least 79%, at least 80%, at least 81%, atleast 82%, atleast 83%, atleast 84%, at least 85%, at least 86%, at least 87%, at least 88%, atleast 89%, atleast 90%, atleast 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even by 100%. Indeed, a suitable increase may be by at least two-fold, at least 5 -fold, at least 10-fold, at least 25-fold, at least 50-fold, or at least 100-fold or more.

[0167] In some cases, SC-Ads suitable for use in the methods of treatment or medical uses of the invention canbe administered by inhalation as a heterologous or homologous vaccination boost to a mammal (e.g., a human) that previously received a vaccine targeting the same pathogen (e.g., a bacterial or a viral pathogen) to produce IgG and IgA antibodies against the immunogen(s) within the mammal starting from about 15 days post administration (or starting from about 16 days post administration, or starting from about 17 days post administration, or starting from about 18 days post administration, or starting from about 19 days post administration, or starting from about 20 days post administration, or starting from about 21 days post administration, or starting from about22 dayspostadministration) and lasting atleast to about 40 days post administration (e.g., lasting at least to about 43 days post administration, lasting at least to about 45 days post administration, lasting at least to about 50 days post administration, lasting at least to about 55 days post administration, lasting at least to about 60 days post administration, lasting at least to about 65 days post administration, lasting at least to about 70 days post administration, lasting at least to about 75 days post administration, lasting at least to about 80 days post administration, lasting at least to about 85 days post administration, lasting at least to about 90 days post administration, lasting at least to about 95 days post administration, lasting at least to about 100 days post administration, lasting atleast to about 105 days post administration, lasting at least to about 110 days post administration, lasting at least to about 115 days post administration, lasting at least to about 120 days post administration, lasting atleast to about 125 days post administration, lasting at least to about 130 days post administration, or lasting at least to about 135 days post administration).

[0168] In some cases, SC-Ads suitable for use in the methods of treatment or medical uses of the invention canbe administered by inhalation as a heterologous or homologous vaccination boost to a mammal (e.g., a human) that previously received a vaccine targeting the same pathogen (e.g., a bacterial or a viral pathogen) to increase a T cell response to the pathogen within the mammal. For example, adenovirus vectors describedherein encoding one or more immunogens can be administered by inhalation as a heterologous or homologous vaccination boost to a mammal that previously received a vaccine targeting the same pathogen (e.g., a bacterial or a viral pathogen) to increase the number of activated T cells (e.g., activated Thl T cells) within the mammal by at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21%, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31%, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least 40%, at least 41 %, at least 42%, at least 43%, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51%, at least 52%, at least 53%, atleast 54%, atleast 55%, atleast 56%, at least 57%, at least 58%, at least 59%, at least 60%, atleast 61%, atleast 62%, atleast 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least

[0169] 71%, at least 72%, at least 73%, at least 74%, atleast 75%, atleast 76%, atleast 77%, at least

[0170] 78%, at least 79%, at least 80%, at least 81%, atleast 82%, atleast 83%, atleast 84%, at least 85%, at least 86%, at least 87%, at least 88%, atleast 89%, atleast 90%, atleast 91%, at least

[0171] 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least

[0172] 99%, or even by 100%. Indeed, a suitable increase may be by at least two-fold, atleast 5 -fold, at least 10-fold, at least 25-fold, at least 50-fold, or at least 100-fold or more.

[0173] As described herein, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention is administered by inhalation as a heterologous or homologous vaccination boost to a mammal (e.g., a human) that previously received a vaccine targeting the same pathogen (e.g., a bacterial or a viral pathogen) to induce IgG antibodies, IgA antibodies, and / or Thl T cell responses (e.g., skewed Thl T cell responses) within the mammal. For example, a SC-Ad engineered to encode a SARS-CoV-2 Spike polypeptide can be administered by inhalation as a heterologous vaccination boost to a human who previously received an mRNA-based or DNA-based SARS-CoV-2 vaccine (e.g., a Pfizer® vaccine such as Comirnaty®, an AstraZeneca® vaccine such as Vaxzevria®, a Moderna® vaccine such as Spikevax®, a Novavax® vaccine such as Nuvaxovid®, and / or Sputnik) to induce IgG antibodies, IgA antibodies, and / or Thl T cell responses against coronaviruses within the human.

[0174] In a suitable embodiment a method of treatment, comprises the administration of said SC-Ad by inhalation is provided as a vaccine prime to a mammal that will receive a further vaccination boost to the pathogen after said administration of said SC-Ad by inhalation. Suitably the further vaccination boost may be administered at least one week, at least two weeks, at least three weeks, at least four weeks, at least five weeks, or at least six weeks, or more, after the administration of the SC-Ad by inhalation. Suitably, the further vaccination boost may be administered atleast 50 days, atleast 60 days, at least 70 days, at least 80 days, atleast 90 days, at least 100 days, at least 110 days, atleast 120 days, at least 130 days, at least 140 days, or at least 150 days after said administration of the SC-Ad by inhalation.

[0175] Suitably in such an embodiment said further vaccination will not comprise administration of said SC-Ad (i.e. a heterologous prime-boost approach, in which the administration of the SC- Ads represents the prime). Alternatively, in a suitable embodiment the further vaccination will comprise administration of said SC-Ad (i.e. a homologous prime-boost approach, in which administration of the SC-Ads is used to provide both the prime and the boost).

[0176] Except for where context requires otherwise, the suggestions set out in the preceding paragraphs under the “Treatment regimes” heading (e.g. those regarding timing between prime and boost, the nature and / or quantification of the immune response induced, and assessment of efficacy) apply equally to embodiments of the methods of treatment or medical uses of the invention employing administration of the SC-Ads as a prime, as to those employing administration of the SC-Ads as a boost.

[0177] Having the ability to perform heterologous or homologous vaccination boosts produce immune responses effectively against pathogens (e.g., a coronavirus) in mammals (e.g., humans) can improve survival and minimize the impact of the infection.

[0178] Pharmaceutical compositions or formulations

[0179] In the third aspect, the invention provides a pharmaceutical composition comprising:

[0180] • a SC-Ad comprising a nucleic acid sequence encoding an immunogen of a pathogen; and

[0181] • excipients suitable for use in administration of said SC-Ad to a mammal by inhalation.

[0182] The SC-Ad(s) incorporated in a pharmaceutical composition of the invention may be in accordance with any of the embodiments disclosed in the present invention. In a suitable embodiment, a pharmaceutical composition of the invention comprises excipients in the form of a liquid formulation comprising one, more, or all of: NaCl, histidine, MgC12, EDTA, polysorbate 80, sucrose, and ethanol.

[0183] In a suitable embodiment, a pharmaceutical composition of the invention comprises excipients in the form of a liquid formulation comprising all of: NaCl, histidine, MgC12, EDTA, polysorbate 80, sucrose, and ethanol.

[0184] A pharmaceutical composition of the invention may be formulated to provide a therapeutically effective amount of a SC-Ad, as considered below in this specification, in a selected number of dosage units (i.e., to provide a therapeutically effective amount of a SC-Ad in one, two, three, four, five, or more incidences of inhalation).

[0185] In some cases, SC-Ads suitable for use in the methods of treatment or medical uses of the invention encoding one or more immunogens (and / or nucleic acid molecules that can encode an SC-Ad suitable for use in the methods of treatment or medical uses of the invention encoding one or more immunogens) can be formulated into a composition (e.g., a pharmaceutical composition such as a vaccine composition) for administration to a mammal. For example, SC-Ads suitable for use in the methods of treatment or medical uses of the invention encoding one or more immunogens (and / or nucleic acid molecules that can encode a SC-Ad suitable for use in the methods of treatment or medical uses of the invention encoding one or more immunogens) can be formulated together with one or more pharmaceutically acceptable carriers (additives), excipients, and / or diluents. Examples of pharmaceutically acceptable carriers, excipients, and diluents that can be used in a composition include, without limitation, sucrose, lactose, starch (e.g., starch glycolate), cellulose, cellulose derivatives (e.g., modified celluloses such as microcrystalline cellulose, and cellulose ethers like hydroxypropyl cellulose (HPC) and cellulose ether hydroxypropyl methylcellulose (HPMC)), xylitol, sorbitol, mannitol, gelatin, polymers (e.g., polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), crosslinked polyvinylpyrrolidone (crospovidone), carboxymethyl cellulose, polyethylene- polyoxypropylene-block polymers, and crosslinked sodium carboxymethyl cellulose (croscarmellose sodium)), titanium oxide, azo dyes, silica gel, fumed silica, talc, magnesium carbonate, vegetable stearin, magnesium stearate, aluminum stearate, stearic acid, antioxidants (e.g., vitamin A, vitamin E, vitamin C, retinyl palmitate, and selenium), citric acid, sodium citrate, parabens (e.g., methyl paraben and propyl paraben), petrolatum, dimethyl sulfoxide, mineral oil, serum proteins (e.g., human serum albumin), glycine, sorbic acid, potassium sorbate, water, salts or electrolytes (e.g., saline, protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, and zinc salts), colloidal silica, magnesium trisilicate, polyacrylates, waxes, wool fat, lecithin, and corn oil. Suitable pharmaceutical formulations depend in part upon the use and the route of administration. Such forms should not prevent the composition or formulation from reaching target cells or from exerting its effect. For example, pharmacological compositions injected into the blood stream should be soluble.

[0186] In a suitable embodiment, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention is formulated as a liquid formulation. Liquid formulations refer to pharmaceutical or chemical products that are formulated as liquids rather than solids or semisolids. These formulations are typically composed of one or more active ingredients dissolved or suspended in a liquid solvent or vehicle, along with other additives such as preservatives or stabilizers. In the context provided herein, SC-Ads suitable for use in the methods of treatment or medical uses of the invention may be dissolved or suspended in a liquid solvent or vehicle.

[0187] The liquid formulation may be formulated together with one or more pharmaceutically acceptable carriers (additives), excipients, and / or diluents as discussed above. In a suitable embodiment, the liquid formulation comprisesNaCl, histidine, MgC12, EDTA, polysorbate 80, sucrose, and ethanol.

[0188] In a suitable embodiment, the SC-Ad is formulated as a solid powder. Solid powder typically refers to a dry, stable form of SC-Ads that are preserved in a solid state. The liquid formulation may be formulated together with one or more pharmaceutically acceptable carriers (additives), excipients, and / or diluents as discussed above.

[0189] In some cases, a composition including SC-Ads suitable for use in the methods of treatment or medical uses of the invention encoding one or more immunogens (and / or nucleic acid molecules that can encode a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can include a plurality of identical SC-Ads that are designed to include one or more immunogens (and / or nucleic acid molecules that can encode a SC-Ad suitable for use in the methods of treatment or medical uses of the invention encoding one or more immunogens).

[0190] In some cases, a composition including SC-Ads suitable for use in the methods of treatment or medical uses of the invention encoding one or more immunogens (and / or nucleic acid molecules that can encode a SC-Ad suitable for use in the methods of treatment or medical uses of the invention encoding one or more immunogens) can include a population of two or more (e.g., two, three, four, five, or more) different adenovirus vectors. For example, a composition can be designed to include two populations of SC-Ads suitable for use in the methods of treatment or medical uses of the invention , with the first population containing one or more coronavirus immunogens (e.g., an amino acid sequence set forth in any one of SEQ ID NOs: 1-4, 42-44, 47, and 48) and the second population containing one or more adjuvant polypeptides (e.g., a GM-CSF polypeptide, an IL-4 polypeptide, an IL-21 polypeptide, a CD40L polypeptide, a 4-1BBL polypeptide, a TGF-P polypeptide, a C difficile TcdA polypeptide, a C. difficile TcdB polypeptide, and / or biologically active fragments thereof). For example, a composition can be designed to include two populations of SC-Ads suitable for use in the methods of treatment or medical uses of the invention , with the first population containing one or more coronavirus immunogens (e.g., an amino acid sequence set forth in any one of SEQ ID NOs: 1-4, 42-44, 47, and 48) and the second population containing a chaff polypeptide (e.g., a fragment of an ACE2 polypeptide).

[0191] In some cases, a composition including SC-Ads suitable for use in the methods of treatment or medical uses of the invention can be designed to include populations of different SC-Ads where each population of SC-Ads of the composition contains a single immunogen. For example, a composition of SC-Ads suitable for use in the methods of treatment or medical uses of the invention can be designed to include a first population of SC-Ads suitable for use in the methods of treatment or medical uses of the invention having a first coronavirus immunogen (e.g., an amino acid sequence set forth in any one of SEQ ID NOs: 1-4, 42-44, 47, and 48) and a second population of SC-Ads suitable for use in the methods of treatment or medical uses of the invention having a second coronavirus immunogen (e.g., an amino acid sequence set forth in any one of SEQ ID NOs: 1 -4, 42-44, 47, and 48). For example, a composition or formulation of SC-Ads suitable for use in the methods of treatment or medical uses of the invention can be designed to include a first population of SC-Ads suitable for use in the methods of treatment or medical uses of the invention having a first coronavirus immunogen (e.g., an amino acid sequence set forth in any one of SEQ ID NOs: 1-4, 42-44, 47, and 48), a second population of SC-Ads suitable for use in the methods of treatment or medical uses of the invention having a second coronavirus immunogen (e.g., an amino acid sequence set forth in any one of SEQ ID NOs: 1 -4, 42-44, 47, and 48), and a third population of SC-Ads suitable for use in the methods of treatment or medical uses of the invention having a third coronavirus immunogen (e.g., an amino acid sequence set forth in any one of SEQ ID NOs: 1-4, 42-44, 47, and 48). In another example, a composition or formulation of SC-Ads suitable for use in the methods of treatment or medical uses ofthe invention can be designedto include a first population of SC-Ads suitable for use in the methods of treatment or medical uses of the invention having a first coronavirus immunogen (e.g., an amino acid sequence setforth in any one of SEQ ID NOs: 1-4, 42-44, 47, and 48), a second population of SC-Ads suitable for use in the methods of treatment or medical uses of the invention having a second coronavirus immunogen (e.g., an amino acid sequence set forth in any one of SEQ ID NOs: 1 -4, 42-44, 47, and 48), and a third population of SC-Ads suitable for use in the methods of treatment or medical uses of the invention an adjuvant polypeptide (e.g., a GM-CSF polypeptide, an IL-4 polypeptide, an IL-21 polypeptide, a CD40L polypeptide, a 4-1BBL polypeptide, aTGF-P polypeptide, a C difficile TcdA polypeptide, a C. difficile TcdB polypeptide, and / or biologically active fragments thereof), a first population of SC-Ads suitable for use in the methods of treatment or medical uses of the invention having a first coronavirus immunogen (e.g., an amino acid sequence set forth in any one of SEQ ID NOs: 1 -4, 42-44, 47, and 48), a second population of SC-Ads suitable for use in the methods of treatment or medical uses of the invention having a second coronavirus immunogen (e.g., an amino acid sequence set forth in any one of SEQ ID NOs: 1 -4, 42-44, 47, and 48), and a third population of SC-Ads suitable for use in the methods of treatment or medical uses of the invention having a chaff polypeptide (e.g., a fragment of an ACE2 polypeptide).

[0192] Therapeutically effective amounts of SC-Ads suitable for use in the methods of treatment or medical uses of the invention

[0193] The methods of treatment or medical uses of the invention involve the provision of therapeutically effective amounts of SC-Ads suitable for use in the methods or uses of the invention. The therapeutically effective amount of a SC-Ad may be Such therapeutically effective amounts of the requisite SC-Ads may be provided in a single incidence of treatment, or may be accumulated over the course of a number of incidences of treatment. SC-Ads suitable for use in the methods or uses of the invention can be administered to a mammal (e.g., a human) in any appropriate amount (e.g., any appropriate dose). Effective amounts can vary depending on the route of administration, the age and general health condition of the subject, excipient usage, the possibility of co-usage with other therapeutic treatments such as use of other agents, and the judgment of the treating physician.

[0194] An effective amount of a composition containing SC-Ads suitable for use in the methods or uses of the invention can be any amount that can induce an immune response in a mammal as described herein without producing significant toxicity to the mammal.

[0195] A therapeutically effective amount in respect of a SC-Ad suitable for use in the methods of treatment or medical uses of the invention, may be defined in terms of the amount of viral particles.

[0196] In a suitable embodiment, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention is administered in a therapeutically effective amount of at least 1 x 103, 1 x 104, 1 x 105, 1 x 106, 1 x 107, 1 x 108, 1 x 109, or 1 x 1010SC-Ad viral particles. For example, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention may be administered in a therapeutically effective amount of at least lxl0A10 viral particles, at least 2xl0A10 viral particles, at least 3x10A10 viral particles, at least viral particles 4x10A10, atleast 5xl0A10 viral particles, atleast 6x10A10 viral particles, atleast 7x10A10 viral particles, atleast 8xl0A10 viral particles, atleast 9x10A10 viral particles, atleast 1x10Al 1 viral particles, atleast l . lxlOAl 1 viral particles, atleast 1.2xlOAl 1 viral particles, atleast 1.3xl0Al 1 viral particles, at least 1.4xlOAl 1 viral particles, at least 1.5xl0Al 1 viral particles, at least 1.6xlOAl 1 viral particles, at least 1 ,7x 10Al 1 viral particles, atleast 1 .8x10Al 1 viral particles, atleast 1 ,9x 10Al 1 viral particles, at least 2xlOAl l viral particles, at least 2.1xlOAl l viral particles, at least 2.2x10Al 1 viral particles, atleast 2.3x10Al 1 viral particles, atleast 2.4x10Al 1 viral particles, at least2.5xlOAl 1, atleast2.6xlOAll viral particles, atleast 2.7xlOAl l viral particles, at least 2.8x 10Al 1 viral particles, atleast 2.9xlOAll viral particles, or at least 3x10Al 1 viral particles.

[0197] In a suitable embodiment, a SC-Ad suitable for use in the methods of treatmentor medical uses of the invention is administered in a therapeutically effective amount of atleast 1 x 103, 1 x 104, 1 x 105, 1 x 106, 1 x 107, 1 x 108, 1 x 109, or 1 x 1010infectious SC-Ad particles. For example, a SC-Ad suitable foruse in the methods of treatment or medical uses of the invention may be administered in a therapeutically effective amount of at least lxlOA8 infectious particles, at least 2x10A8 infectious particles, at least 3x10A8 infectious particles, at least infectious particles 4xlOA8, at least 5xlOA8 infectious particles, at least 6xlOA8 infectious particles, at least 7x10A8 infectious particles, at least 8x10A8 infectious particles, at least 9x10A8 infectious particles, at least lxlOA9 infectious particles, at least l . lxlOA9 infectious particles, at least 1.2xlOA9 infectious particles, at least 1.3xlOA9 infectious particles, at least 1 .4x10A9 infectious particles, atleast 1 .5x10A9 infectious particles, at least 1.6x10A9 infectious particles, at least 1.7xlOA9 infectious particles, at least 1.8xlOA9 infectious particles, at least 1.9xlOA9 infectious particles, at least 2xlOA9 infectious particles, at least 2.1x10A9 infectious particles, at least 2.2xlOA9 infectious particles, at least 2.3x10A9 infectious particles, at least 2.4xlOA9 infectious particles, atleast 2.5xlOA9, atleast2.6xlOA9 infectious particles, atleast 2.7xlOA9 infectious particles, at least 2.8xlOA9 infectious particles, at least 2.9xlOA9 infectious particles, or at least 3x10A9 infectious particles.

[0198] In a suitable embodiment, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention is administered in a therapeutically effective amount of between 1 x 105and 3 x 1011SC-Ad viral particles. For example, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention may be administered in a therapeutically effective amount of between 2xl0A10 viral particles and 2.9xlOAl 1 viral particles, 3xl0A10 viral particles and 2.8xlOAl 1 viral particles, 4xl0A10 viral particles and2.7xlOAl 1 viral particles, 5xl0A10 viral particles and 2.6xlOAl 1 viral particles, 6xl0A10 viral particles and 2.5xlOAl 1 viral particles, 7xl0A10 viral particles and 2.4xlOAl 1 viral particles, 8xl0A10 viral particles and 2.3xlOAll viral particles, 9xl0A10 viral particles and 2.2x10Al 1 viral particles, lxlOAl 1 viral particles and 2.1xlOAll viral particles, l . lxlOAl 1 viral particles and 2.0xl0Al l viral particles, 1.2xlOAl l viral particles and 1.9xlOAl l viral particles, 1.3xlOAl l viral particles and 1 .8x10Al 1 viral particles, 1.4xlOAl 1 viral particles and 1 ,7xlOAll viral particles, or 1.5xlOAl l viral particles and 1.6xlOAl 1 viral particles.

[0199] In a suitable embodiment, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention is administered in a therapeutically effective amount of between 1 x 108and 3 x 109infectious SC-Ad particles. For example, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention may be administered in a therapeutically effective amount of between 2xlOA8 infectious particles and 2.9xlOA9 infectious particles, 3xlOA8 infectious particles and 2.8x10A9 infectious particles, 4xlOA8 infectious particles and2.7x!0A9 infectious particles, 5xlOA8 infectious particles and 2.6xlOA9 infectious particles, 6xlOA8 infectious particles and 2.5x10A9 infectious particles, 7xlOA8 infectious particles and2.4xlOA9 infectious particles, 8xlOA8 infectious particles and 2.3xlOA9 infectious particles, 9xlOA8 infectious particles and 2.2x10A9 infectious particles, lx 10A9 infectious particles and2.1xlOA9 infectious particles, 1 .1x10A9 infectious particles and 2.0xl0A9 infectious particles, 1 ^xlC ^ infectious particles and 1.9xlOA9 infectious particles, 1.3xlOA9 infectious particles and 1 ,8xlOA9 infectious particles, 1.4xlOA9 infectious particles and 1 .7x10A9 infectious particles, or 1.5xlOA9 infectious particles and 1.6xlOA9 infectious particles.

[0200] The effective amount can remain constant or can be adjusted as a sliding scale or variable dose depending on the mammal’s response to treatment. Various factors can influence the actual effective amount used for a particular application. For example, the frequency of administration, duration of treatment, use of multiple treatment agents, and / or route of administration may require an increase or decrease in the actual effective amount administered.

[0201] Manufacture of SC-Ad for use in method of treatment or medical uses of the invention

[0202] A SC-Ad to b e used in the methods of treatment or medical uses of the invention can b e derived from any suitable adenovirus. An adenovirus used to create a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can be any appropriate serotype (e.g., Adi - Ad57). By way of example, suitable adenoviruses that can be used to make a SC-Ad for use in the methods oftreatment or medical uses of the invention include, without limitation, Ad5 adenoviruses, Ad6 adenoviruses, ChAdOxl, and ChAdOx2.

[0203] It will be appreciated that irrespective of the adenovirus used to create a SC-Ad for use in the invention (the “starting virus”), the SC-Ad itself will always only be able to undergo a single cycle of replication.

[0204] In some cases, the starting adenovirus used to create a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can be a replication defective adenovirus. In some cases, the starting adenovirus can be a replication competent adenovirus. If a replication competent adenovirus was used as the starting virus to create a SC-Ad of the invention, the resulting SC-Ad will still only be able to undergo a single cycle of replication. An adenovirus used to create a SC-Ad suitable for use in the methods of treatment or medical uses of the invention may be capable of infecting a human cell (i.e., the starting virus can be a human adenovirus).

[0205] In some cases, an adenovirus used to create a SC-Ad suitable for use in the methods of treatment or medical uses of the invention may be capable of infection a non-human cell such as chimpanzee cells (i.e., the starting virus can be a non-human adenovirus).

[0206] An adenovirus for use in treatment of a human subject may be created from a human starting adenovirus.

[0207] Structure of SC-Ads for use in method of treatment or medical uses of the invention

[0208] As noted above, SC-Ads may lack one or more functional genes essential for viral replication. Suitably an SC-Ad lacks one or more functional gene selected from the group consisting of genes involved in DNA replication; genes involved in capsid formation; and genes involved in virion assembly. The lack of one or more such genes means that the adenovirus is unable to complete a full replication cycle and produce infectious virions.

[0209] Suitably one or more of the genes lacking from the SC-Ad are deleted or inactivated.

[0210] In some embodiments, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention has a genome which lacks all or a portion of at least one early or late gene productencoding sequence. A suitable SC-Ad may lack all or a portion of at least one adenovirus nucleic acid sequence selected from the group consisting of: fiber protein-encoding sequence; V protein-encoding sequence; hexon-encoding sequence; pentonbase-encoding sequence (also referred to as a pill-encoding sequence); VA RNA-encoding sequence; and pllla proteinencoding sequence (also referred to as a minor capsid protein-encoding sequence). Examples of nucleic acid sequences that encode adenoviral polypeptides include, without limitation, those set forth in GenBank gi numbers 209842, 58478, or 2935210, and / or annotated in GenBank accession numbers M73260, X17016, or AF030154.

[0211] In some cases, a deletion of all or a portion of the nucleic acid can be engineered into a nucleic acid such that the SC-Ad does not encode one or more early or late gene product-encoding sequences. Suitably, a deletion of all or a portion of the nucleic acid encoding one or more of the following polypeptides can be engineered into a nucleic acid encoding an adenovirus such that the SC- Ad does not encode one or more full-length adenovirus polypeptide, or one or more fully functional version of an adenovirus polypeptide, selected from the group consisting of fiber protein-encoding sequence; V protein-encoding sequence; hexon-encoding sequence; penton base-encoding sequence; VA RNA-encoding sequence; and pllla protein-encoding sequence.

[0212] Suitable deletions in nucleic acid sequences, such as those considered above, can be of any length that results in the deletion of one or more encoded amino acids and in a reduction or elimination of the normal function of that polypeptide. For example, portions of a nucleic acid sequence of an adenovirus can be removed such that the otherwise encoded polypeptide lacks 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, or more amino acid residues and lacks its normal activity.

[0213] The portion or portions to be deleted can be removed from any location along the length of the sequence. For example, a portion of an adenovirus nucleic acid sequence can be removed at the 5 ’ end, the 3 ’ end, or an internal region of an adenovirus nucleic acid such as a fiber proteinencoding sequence, V protein-encoding sequence, hexon-encoding sequence, penton baseencoding sequence, VA RNA-encoding sequence, pllla protein-encoding sequence, or other early or late gene product-encoding sequences.

[0214] Suitably, a SC-Ad for use in the methods of treatment or medical uses of the invention can be as described elsewhere (see, e.g., Matchett etal., J. of Virol., 2019 93(10):e02016-18 (2019); and International PCT Patent Application Publication No. WO 2009 / 111738).

[0215] A SC-Ad suitable for use in the methods of treatment or medical uses of the invention can include a nucleic acid sequence encoding any appropriate immunogen (e.g., a nucleic acid that drives expression of any appropriate immunogen). In some cases, an immunogen can be an antigen. An immunogen can be a full-length immunogenic polypeptide or a portion thereof (e.g., can be derived from an immunogenic polypeptide).

[0216] The immunogen can be from any type of pathogen (e.g., a virus, a bacterium, a protozoan, a prion, a viroid, or a fungus). In some cases, an immunogen can be a polypeptide expressed by a virus (e.g., a viral polypeptide). For example, an immunogen can be a polypeptide expressed by a coronavirus (e.g., a beta-coronavirus). Suitable examples of viral sources of immunogens include, without limitation, SARS-CoV, HCoVNL63, HKU1, MERS-CoV, SARS-CoV-2, HIV-1, hepatitis B virus, hepatitis C virus, hepatitis D virus, hepatitis E virus, influenza, Ebolavirus, Chikungunya virus, Zika virus, cytomegalovirus, West Nile virus, and those described in Table 3-1 of “Learning from SARS: Preparing for the Next Disease Outbreak: Workshop Summary.” Institute of Medicine (US) Forum on Microbial Threats; Knobler S, Mahmoud A, Lemon S, et al., editors. Washington (DC): National Academies Press (US); 2004.

[0217] In some cases, an immunogen can be a polypeptide expressed by a bacterium (e.g., a bacterial polypeptide), or a polypeptide derived from such a bacterial polypeptide. Suitable examples of bacterial sources of immunogens include, without limitation, Clostridium (e.g., C. difficile), Staphylococcus aureus (e.g. methicillin-resistant S. aureus), Campylobacter (e.g. Campylobacter jejuni), Mycobacteria (e.g. M. tuberculosis), and Borrelia (B. burgdorferi .

[0218] Examples of immunogens that may be encoded by a nucleic acid incorporated in a SC-Ad suitable for use in the methods of treatment or medical uses of the invention include, without limitation, C. difficile Toxin A (TcdA) polypeptides, C. difficile Toxin B (TcdB) polypeptides, coronavirus Spike polypeptides, the amino acid sequence set forth in SEQ ID NO:1, coronavirus nucleoproteins, coronavirus membrane proteins, coronavirus envelope proteins, and coronavirus non-structural proteins (e.g., coronavirus non-structural proteins 1-16).

[0219] In a suitable example, an immunogen that may be encoded by a nucleic acid incorporated in a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can have, or can be encoded by, a sequence set forth in, for example, National Center for Biotechnology Information (NCB I) Accession Nos: MN938384 and AY772062.

[0220] In some cases, an immunogen is from an opportunistic pathogen.

[0221] In some cases, an immunogen is from a respiratory pathogen. A suitable immunogen can be from any respiratory pathogen of interest. For the purposes of the present disclosure, a respiratory pathogen refers to any microorganism capable of causing infections primarily affecting the respiratory tract. These pathogens include viruses, bacteria, fungi, and other infectious agents that can infect the upper respiratory tract (e.g., nose, throat, sinuses) and / or the lower respiratory tract (e.g., bronchi, lungs). A suitable respiratory pathogen may be an opportunistic pathogen.

[0222] In a suitable embodiment, an immunogen encoded by a nucleic acid incorporated in a SC-Ad suitable for use in the methods of treatment or medical uses of the invention may be selected from the group consisting of: a SARS-CoV-2 immunogen, a SARS-CoV immunogen, a HCoV NL63 immunogen, a HKU1 immunogen, a MERS-CoV immunogen, an influenza immunogen, a respiratory syncytial virus (RSV) immunogen, a metapneumovirus (MPV) immunogen, a rhinovirus immunogen, a bocavirus immunogen, a parainfluenza virus (PIV) immunogen, a Streptococcus pneumoniae immunogen, a Bordetella pertussis immunogen, a Haemophilus influenzae immunogen, a Mycobacterium tuberculosis immunogen, and a Klebsiella pneumonia immunogen.

[0223] A nucleic acid sequence encoding a SC-Ad with a spike protein immunogen is set out in SEQ ID NO:26.

[0224] An immunogen that may be encoded in a SC-Ad suitable for use in the methods of treatment or medical uses of the invention may be from an opportunistic pathogen selected from the group consisting of: CMV, cryptococcus, aspergillus, histoplasma, nocardia, mycobacterium avium, Legionella pneumophila, Candida species, toxoplasma pneumonia, non-tuberculous mycobacteria, and invasive fungal species (such as mucormycetes).

[0225] An immunogen that may be encoded in a SC-Ad suitable for use in the methods of treatment or medical uses of the invention may be from an opportunistic pathogen causing an opportunistic infection selected from the group consisting of: pneumocystis pneumonia (PCP), Cytomegalovirus (CMV) pneumonia, Cryptococcal pneumonia, Aspergillosis, Histoplasmosis, Nocardiosis, Mycobacterium avium complex (MAC) infection, Pneumonia caused by atypical pathogens (such as Legionella pneumophila), Fungal pneumonia caused by Candida species, Toxoplasmosis pneumonia, Respiratory infections caused by non-tuberculous mycobacteria (NTM), and invasive fungal infection (such as mucormycosis). A nucleic acid sequence encoding an immunogen may be modified to remove a naturally occurring ER retention sequence, such that the encoded immunogen, when expressed, lacks a naturally occurring ER retention sequence.

[0226] An immunogen can be designed to extend from cells infected with the SC- Ad vector into the extracellular space. For example, a nucleic acid sequence encoding an immunogen may be modified to encode an immunogen including an ectodomain of an immunogenic polypeptide.

[0227] In some cases, an immunogen can bind (e.g., can be designed to bind) to viral receptor (e.g., an ACE2 polypeptide). For example, an immunogen can include a receptor binding domain of an immunogenic polypeptide.

[0228] In some cases, an immunogen can include (e.g., can be a fusion polypeptide of) two or more immunogenic polypeptides described herein. For example, an immunogen can include a first immunogenic polypeptide and a second immunogenic polypeptide. In some embodiments, a first immunogenic polypeptide and a second immunogenic polypeptide are different polypeptides. For example, an immunogen can include a tcdA polypeptide and a tcdB polypeptide (e.g., a tcdA / B fusion polypeptide). Anucleic acid sequence encoding a SC-Ad with a TcdA / B immunogen is set out in SEQ ID NO:27. In some embodiments, a first immunogenic polypeptide and a second immunogenic polypeptide are the same polypeptide. For example, an immunogen can include a tcdB polypeptide and a tcdB polypeptide (e.g., a tcdB / B fusion polypeptide).

[0229] When an immunogen includes a first immunogenic polypeptide and a second immunogenic polypeptide, the first immunogenic polypeptide can be derived from a first pathogen, and the second immunogenic polypeptide can be derived from a second pathogen. The first pathogen and the second pathogen can be the same pathogen or different pathogens.

[0230] When the first pathogen andthe second pathogen are the same pathogen, the first immunogenic polypeptide and the second immunogenic polypeptide can be derived from the different strains of that pathogen. For example, a first immunogenic polypeptide and a second immunogenic polypeptide can be derived from different strains of the same bacterium (e.g., C. difficile). Examples of immunogens derived from immunogenic polypeptides that can be used as described herein include, without limitation, the amino acid sequence set forth in SEQ ID NO:2, the amino acid sequence set forth in SEQ ID N0:3, the amino acid sequence set forth in SEQ ID NO:4 , the amino acid sequence set forth in SEQ ID NO: 11 , the amino acid sequence set forth in SEQ ID NO: 12 , the amino acid sequence set forth in SEQ ID NO: 13 , the amino acid sequence set forth in SEQ ID NO: 14, the amino acid sequence set forth in SEQ ID N0:15, the amino acid sequence set forth in SEQ ID NO: 16, the amino acid sequence set forth in SEQ ID NO: 17, the amino acid sequence set forth in SEQ ID NO: 18, the amino acid sequence set forth in SEQ ID NO: 19, the amino acid sequence set forth in SEQ ID NO:42, the amino acid sequence set forth in SEQ ID NO:43, the amino acid sequence set forth in SEQ ID NO:44, the amino acid sequence set forth in SEQ ID NO:45, the amino acid sequence set forth in SEQ ID NO:46 , the amino acid sequence set forth in SEQ ID NO:47, and the amino acid sequence set forth in SEQ ID NON 8.

[0231] In some cases, an immunogen encoded by a nucleic acid sequence present in a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can be a variant of a wildtype immunogen. For example, a suitable immunogen may be a variant of a coronavirus Spike polypeptide (e.g., a SARS-CoV-2 Spike polypeptide) comprising or consisting essentially of an amino acid sequence set forth in any one of SEQ ID NOs:l-4 with one or more (e.g., one, two, three, four, five, six, seven, eight, nine, ten, or more) amino acid deletions, additions, substitutions, or combinations thereof.

[0232] Examples of amino acid deletions that can be present in a variant of a coronavirus Spike polypeptide (e.g., as numbered in any one of SEQ ID NOs:l-4) include, without limitation, a deletion ofresidues 69-70, a deletion of residue 144, a deletion of residues 156-157, a deletion of residues 241-243, and a deletion of residues 246-252.

[0233] Examples of amino acid substitutions that can be present in a variant of a coronavirus Spike polypeptide (e.g., as numbered in any one of SEQ ID NOs:l-4), include, without limitation, a L5F amino acid substitution, a S13I amino acid substitution, a L18F amino acid substitution, a T19R amino acid substitution, a T20N amino acid substitution, aP26S amino acid substitution, a G75I amino acid substitution, a A67V amino acid substitution, a V70F amino acid substitution, a T76I amino acid substitution, a D80Aamino acid substitution, a D80G amino acid substitution, a T95I amino acid substitution, a D138Y amino acid substitution, a G142D amino acid substitution, a W152C amino acid substitution, a El 54K amino acid substitution, a F157S amino acid substitution, a R158G amino acid substitution, a R190S amino acid substitution, a D215G amino acid substitution, a A222V amino acid substitution, a D253G amino acid substitution, a W258L amino acid substitution, a K417N amino acid substitution, a K417T amino acid substitution, a L452R amino acid substitution, a L452Q amino acid substitution, a Y453F amino acid substitution, a S477N amino acid substitution, a T478K amino acid substitution, a E484Q amino acid substitution, a E484K amino acid substitution, a F490S amino acid substitution, a E484K amino acid substitution, a S494P amino acid substitution, a N501Y amino acid substitution, a A570D amino acid substitution, a D614G amino acid substitution, a H655 Y amino acid substitution, a Q677H amino acid substitution, a P681H amino acid substitution, a P681R amino acid substitution, a A701V amino acid substitution, a T7161 amino acid substitution, a T859N amino acid substitution, a F888Lamino substitution, a D950N amino acid substitution, a Q957R amino acid substitution, a S982A amino acid substitution, a K986P amino acid substitution, a V987P amino acid substitution, a T1027I amino acid substitution, a Q1071H amino acid substitution, a D1118H amino acid substitution, and a KI 19 IN amino acid substitution. For example, a variant of a coronavirus Spike polypeptide can include a K986P amino acid substitution and a V987P amino acid substitution (e.g., a PP substitution).

[0234] In some cases, a variant of a coronavirus Spike polypeptide can be a gamma mink variant of a coronavirus Spike polypeptide.

[0235] In some cases, a variant of a coronavirus Spike polypeptide can be a delta variant of a coronavirus Spike polypeptide. In some cases, a variant of a coronavirus Spike polypeptide can be a lambda variant of a coronavirus Spike polypeptide. In some cases, a variant of a coronavirus Spike polypeptide can be an epsilon variant of a coronavirus Spike polypeptide. In some cases, a variant of a coronavirus Spike polypeptide can be a delta plus variant of a coronavirus Spike polypeptide.

[0236] In some cases, an immunogen encoded by a nucleic acid sequence present in a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can have an amino acid sequence with at least 85% sequence identity (e.g., at least 88% sequence identity, at least 89% sequence identity, at least 90% sequence identity, at least 91 % sequence identity, at least 92% sequence identity, at least 93% sequence identity, at least 94% sequence identity, at least 95% sequence identity, at least 96% sequence identity, at least 97% sequence identity, at least 98% sequence identity, or at least 99% sequence identity) to the amino acid sequence set forth in any one of SEQ ID NOs:l-4.

[0237] The skilled person will recognise that there are multiple common techniques that may be used to calculate percentage identity. Suitable techniques that may be used to calculate percentage identity include alignment of the sequences using various algorithms available for this purpose, such as BLAST (Basic Local Alignment Search Tool), ClustalW, or specialized protein alignment software like MUSCLE or AFFT followed by counting the matching positions, determining the total aligned positions and calculating the percentage identity using the following formula: Percentage Identity = (Number of Matching Positions / Total Aligned Positions) x 100.

[0238] In some cases, an immunogen encoded by a nucleic acid sequence present in a SC-Ad suitable for use in the methods of treatment or medical uses of the invention may be a coronavirus Spike polypeptide variant containing the entire amino acid sequence set forth in any one of SEQ ID NOs:l-4, exceptthatthe amino acid sequence contains from one to ten (e.g., one to nine, two to nine, one to eight, two to eight, one to seven, one to six, one to five, one to four, one to three, two, or one) amino acid additions, deletions, substitutions, or combinations thereof, provided that the coronavirus Spike polypeptide variant has the ability to induce an immune response against a coronavirus within a mammal (e.g., a human). In some cases, an immunogen encoded by a nucleic acid sequence present in a SC-Ad suitable for use in the methods of treatment or medical uses of the invention may be a coronavirus Spike polypeptide variant consisting essentially of the amino acid sequence set forth in any one of SEQ ID NOs:l-4 exceptthatthe amino acid sequence contains one, two, three, four, or five amino acid residues preceding the articulated sequence ofthe sequence identifier (e.g., SEQ IDNO: 1), and / orhas one, two, three, four, or five amino acid residues following the articulated sequence of the sequence identifier (e.g., SEQ ID NO: 1), provided that the coronavirus Spike polypeptide has the ability to induce an immune response against a coronavirus within a mammal (e.g., a human).

[0239] In some cases, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can include nucleic acid sequence encoding two or more (e.g., two, three, four, five, six, seven, eight, nine, ten, or more) immunogens from different immunogenic polypeptides. For example, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention and can include nucleic acid sequence encoding an immunogen derived from a second pathogen (e.g., an immunogen derived from an immunogenic polypeptide expressed by a pathogen other than SARS-CoV-2). In some cases, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention that includes a nucleic acid sequence encoding two or more immunogens derived from an immunogenic polypeptide expressed by different pathogens can include nucleic acid sequence encoding a polypeptide comprising, consisting of, or consisting essentially of the amino acid sequence set forth in any one of SEQ ID NOs:l-4 and can include nucleic acid sequence encoding a polypeptide comprising, consisting of, or consisting essentially of the amino acid sequence set forth in any one of SEQ ID NOs:20-21. When a SC-Ad suitable for use in the methods of treatment or medical uses of the invention includes nucleic acid sequence encoding two or more immunogens from immunogenic polypeptides expressed by different pathogens, the SC-Ad can be used to induce an immune response against two or more pathogens.

[0240] In some cases, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can include nucleic acid sequence encoding two or more (e.g., two, three, four, five, six, seven, eight, nine, ten, or more) immunogens from the same immunogenic polypeptide and / or the same pathogen. For example, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can include nucleic acid sequence encoding two or more immunogens derived from the same pathogen. In some cases, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention that includes a nucleic acid sequence encoding two or more immunogens derived from an immunogenic polypeptide expressed by influenza can include nucleic acid sequence encoding a polypeptide comprising, consisting of, or consisting essentially of the amino acid sequence setforth in SEQ ID NO:20 and can include nucleic acid sequence encoding a polypeptide comprising, consisting of, or consisting essentially of the amino acid sequence set forth in SEQ ID NO:21.

[0241] In some cases, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention that includes a nucleic acid sequence encoding one or more (e.g., one, two, three, four, five, six, seven, eight, nine, ten, or more) immunogens also can include one or more regulatory sequences (e.g., an enhancer or a promoter sequence such as a constitutive, inducible, and / or tissue-specific promoter sequence) to drive transcription of the immunogen(s). Examples of enhancers and promoters that can be used to drive expression of a nucleic acid sequence encoding one or more immunogens of a SC-Ad suitable for use in the methods of treatment or medical uses of the invention include, without limitation, a CMV enhancer sequence, a CMV promoter sequence, a CAG enhancer sequence, a CAG promoter sequence, a RS V enhancer sequence, a RS V promoter sequence, a Efl alpha enhancer sequence, a Efl alpha promoter sequence, a ubiquitin enhancer sequence, a ubiquitin promoter sequence, adenovirus enhancer sequences, and adenovirus promoter sequences. Any appropriate method can be used to detect expression of an immunogen from adenovirus vector infected cells. For example, antibodies that recognize an immunogencanbeusedto detectthepresenceor absence of the immunogen.

[0242] SC-Ads for use in method of treatment or medical uses of the invention encoding an adjuvant polypeptide

[0243] In some cases, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can include a nucleic acid sequence encoding one or more (e.g., one, two, three, four, five, six, seven, eight, nine, ten, or more) adjuvant polypeptides (e.g., nucleic acid that drives expression of one or more adjuvant polypeptides).

[0244] For example, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can include a nucleic acid sequence encoding one or more polypeptides that can enhance an immune response within a mammal. In some cases, an adjuvant polypeptide can be a cytokine. In some cases, an adjuvant polypeptide can be an immune stimulator. In some cases, an adjuvant polypeptide can be a toxin. In some cases, an adjuvant polypeptide can accelerate a systemic T cell response against a pathogen present within a mammal. In some cases, an adjuvant polypeptide can increase a concentration of antibodies against a pathogen at a site where the pathogen can enter a mammal’s body. For example, an adjuvant polypeptide can increase a concentration of antibodies against a virus (e.g., a coronavirus) at a mucosal site where the virus can enter a mammal’s body.

[0245] Examples of adjuvant polypeptides that can be encoded by a nucleic acid sequence present in a SC-Ad encoding one or more immunogens, suitable for use in a method of treatment or medical uses of the invention described herein include, without limitation, those selected from the group consisting of: granulocyte-macrophage colony -stimulating factor (GM-CSF) polypeptides, interleukin 4 (IL-4) polypeptides, interleukin 21 (IL-21) polypeptides, CD40 ligand (CD40L) polypeptides, 4-1BB ligand (4-1BBL) polypeptides, transforming growth factor beta (TGF-P) polypeptides, C. difficile toxin polypeptides (e.g., C. difficile TcdA polypeptides (see, e.g., SEQ ID NO:22), C. difficile TcdB polypeptides (see, e.g., SEQ ID NO:23), and / or the amino acid sequence set forth in SEQ ID NO: 10), and influenza polypeptides (e.g. N polypeptides, H polypeptides, M polypeptides, the amino acid sequence set forth in SEQ ID NO:20, and / or the amino acid sequence set forth in SEQ ID NO:21).

[0246] In some cases, an adjuvant polypeptide (e.g., SEQ ID NO:22) can be preceded by an AAT secretory sequence (e.g, MPSSVSWGILLLAGLCCLVPVSLAEDP; SEQ ID NO:28). In some cases, a nucleic acid sequence encoding an adjuvant polypeptide (e.g., SEQ ID NO:23) can be preceded by a cleavage site such as a synthetic furin cleavage site (e.g., RGRRSRGRRS; SEQ ID NO:29). Examples of nucleic acid sequences that can encoding an adjuvant polypeptide described herein include, without limitation, the nucleic acid sequence set forth in SEQ ID NO:24) and the nucleic acid sequence set forth in SEQ ID NO:25).

[0247] In some cases, a nucleic acid sequence encoding an adjuvantpolypeptide (e.g., SEQ IDNO:24) can be preceded by an AAT secretory sequence (e.g, ATGCCTTCATCCGTGTCATGGGGAATCCTGCTGCTGGCTGGACTGTGCTGTCTGGT GCCTGTCTCACTGGCCGAGGACCCT; SEQ ID NO:40). In some cases, a nucleic acid sequence encoding an adjuvant polypeptide (e.g, SEQ ID NO:25) can be preceded by a cleavage site such as a synthetic furin cleavage site (e.g, AGAGGACGGAGATCAAGAGGAAGGCGCAGC; SEQ ID NO:41).

[0248] An adjuvant polypeptide can be a full-length polypeptide or a fragment of an adjuvant polypeptide described herein, provided that the fragment has the ability to enhance an immune response (e.g, a biologically active fragment). In some cases, an adjuvantpolypeptide can be as described elsewhere (see, e.g, Matchett et al. Vaccines, 8(1 ):64 (2020)).

[0249] In some cases, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can encode (e.g, can be designed to encode) a polypeptide that includes an immunogen fused to an adjuvant polypeptide. For example, a nucleic acid sequence encoding an immunogen can be fused to a nucleic acid sequence encoding an adjuvant polypeptide (e.g, such that the encoded immunogen is fused to the encoded adjuvant polypeptide). An example of an immunogen fused to an adjuvant polypeptide that can be encoded by an adenovirus vectors encoding one or more immunogens described herein include, without limitation, the amino acid sequence set forth in SEQ ID NO:5.

[0250] SC-Ads for use in method of treatment or medical uses of the invention encoding a chaff polypeptide

[0251] In some cases, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can include a nucleic acid sequence encoding one or more (e.g., one, two, three, four, five, six, seven, eight, nine, ten, or more) chaff polypeptides (e.g., nucleic acid that drives expression of one or more chaff polypeptides). A chaff polypeptide can be a full-length polypeptide or a fragment thereof provided that it reduces the rate of entry or inhibits entry of a pathogen into a cell within a mammal.

[0252] In some cases, a chaff polypeptide can be a soluble polypeptide. For example, a soluble chaff polypeptide can be a full-length chaff polypeptide or a fragment of a chaff polypeptide that lacks a transmembrane domain. For example, a soluble chaff polypeptide can include an ectodomain of a chaff polypeptide. In some cases, a chaff polypeptide can target (e.g., target and bind to) a particular pathogen (e.g., a virus such as a coronavirus) to reduce the rate of entry or inhibit entry of the pathogen into a cell within a mammal. In some cases, a chaff polypeptide can target (e.g., target and bind to) two, three, four, five, six, or more different pathogens. In some cases, a chaff polypeptide can include one or more mutations (e.g., inactivating mutations).

[0253] Examples of chaff polypeptides that can be encoded by a SC-Ad for use in method of treatment or medical uses of the invention encoding one or more immunogens described herein include, without limitation, full-length ACE2 polypeptides and fragments thereof, full-length CD13 polypeptides and fragments thereof, full-length CEACAM1 polypeptides and fragments thereof, full-length sialydated polypeptides and fragments thereof, full-length CD46 polypeptides and fragments thereof, full-length nestin polypeptides and fragments thereof, the amino acid sequence set forth in SEQ ID NO:8 (see, e.g., ), and the amino acid sequence set forth in SEQ ID NO:9.

[0254] In some cases, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can encode (e.g., can be designed to encode) a polypeptide that includes an immunogen fused to a chaff polypeptide. For example, a nucleic acid sequence encoding an immunogen can include a nucleic acid sequence encoding a chaff polypeptide (e.g., such that the encoded immunogen is fused to the encoded chaff polypeptide).

[0255] In some cases, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can include a nucleic acid sequence encoding one or more (e.g., one, two, three, four, five, six, seven, eight, nine, ten, or more) marker polypeptides (e.g., nucleic acid that drives expression of one or more marker polypeptides). Examples of marker polypeptides that can be encoded by a SC-Ad suitable for use in the methods of treatment or medical uses of the invention encoding one or more immunogens described herein include, without limitation, fluorescent polypeptides (e.g., GFP, RFP, CFP, and YFP), streptavidin polypeptides, Cre recombinase polypeptides, Cas polypeptides, luciferase polypeptides, betagalactosidase polypeptides, and sodium iodide symporter polypeptides.

[0256] In some cases, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can include a nucleic acid sequence encoding one or more (e.g., one, two, three, four, five, six, seven, eight, nine, ten, or more) polypeptides that can form a multimer (e.g., nucleic acid that drives expression of one or more polypeptides that can form a multimer). Examples of polypeptides that can form a multimer that can be encoded by a SC-Ad for use in method of treatment or medical uses of the invention encoding one or more immunogens described herein include, without limitation, immunoglobulin constant region polypeptides (e.g., an Ig polypeptide), streptavidin polypeptides, and sigma coil polypeptides.

[0257] In some cases, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can encode (e.g., can be designed to encode) a polypeptide that includes an immunogen fused to a polypeptide that can form a multimer. For example, a nucleic acid sequence encoding an immunogen can include a nucleic acid sequence encoding a polypeptide that can form a multimer (e.g., such that the encoded immunogen is fused to the encoded polypeptide that can form a multimer). Examples of immunogens fused to a polypeptide that can form a multimer that can be encoded by a SC-Ad for use in method oftreatment or medical uses of the invention encoding one or more immunogens described herein include, without limitation, the amino acid sequence set forth in SEQ ID NO:5, the amino acid sequence set forth in SEQ ID NO:6, and the amino acid sequence set forth in SEQ ID NO:7. Sequence structure of SC-Ads suitable for use in the methods of treatment or medical uses of the invention

[0258] In some cases, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can have a genome that is at least 85% percent identical (e.g., at least 88% sequence identical, at least 90% sequence identical, at least 93% sequence identical, at least 95% sequence identical, at least 97% sequence identical, at least 98% sequence identical, or at least 99% sequence identical) to a sequence setforth in any one of SEQIDNOs:28-39. For example, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can have a genome that comprising, consisting of, or consisting essentially of the nucleic acid sequence set forth in SEQ ID NO:28. For example, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can have a genome that comprising, consisting of, or consisting essentially of the nucleic acid sequence set forth in SEQ ID NO:29. For example, a SC-Ad suitable for use in the methods oftreatment or medical uses of the invention can have a genome that comprising, consisting of, or consisting essentially of the nucleic acid sequence setforth in SEQ ID NO:30. For example, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can have a genome that comprising, consisting of, or consisting essentially of the nucleic acid sequence set forth in SEQ ID NO:31 . For example, a SC-Ad suitable for use in the methods oftreatment or medical uses of the invention can have a genome that comprising, consisting of, or consisting essentially of the nucleic acid sequence setforth in SEQ ID NO:32. For example, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can have a genome that comprising, consisting of, or consisting essentially of the nucleic acid sequence set forth in SEQ ID NO:33. For example, SC-Ad suitable for use in the methods oftreatment or medical uses of the invention can have a genome that comprising, consisting of, or consisting essentially of the nucleic acid sequence setforth in SEQ ID NO:34 . For example, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can have a genome that comprising, consisting of, or consisting essentially of the nucleic acid sequence set forth in SEQ ID NO:35. For example, a SC-Ad suitable for use in the methods oftreatment or medical uses of the invention can have a genome that comprising, consisting of, or consisting essentially of the nucleic acid sequence setforth in SEQ ID NO:36 . For example, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can have a genome that comprising, consisting of, or consisting essentially of the nucleic acid sequence set forth in SEQ ID NO:37. For example, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can have a genome that comprising, consisting of, or consisting essentially of the nucleic acid sequence set forth in SEQ ID NO:38 . For example, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can have a genome that comprising, consisting of, or consisting essentially of the nucleic acid sequence set forth in SEQ ID NO:39.

[0259] In some cases, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can have a genome that includes one or more of the coding regions set forth in any one of SEQ ID NOs:28-39. For example, a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can be designed to have a genome where all the encoded polypeptides of the SC-Ad suitable for use in the methods of treatment or medical uses of the invention have the same amino acid sequence as those polypeptides encoded by the nucleic acid set forth in any one of SEQ ID NOs:28-39.

[0260] This document also provides nucleic acid molecules that can encode a SC-Ad suitable for use in the methods of treatment or medical uses of the invention. The term “nucleic acid” as used herein encompasses both RNAandDNA, including cDNA, genomic DNA, and synthetic (e.g., chemically synthesized) DNA. A nucleic acid can be double-stranded or single-stranded. A single-stranded nucleic acid can be the sense strand or the antisense strand. In addition, a nucleic acid can be circular or linear.

[0261] In a suitable embodiment, cells (e.g., cell lines) may contain SC-Ads suitable for use in the methods of treatment or medical uses of the invention. In cases where a SC-Ad suitable for use in the methods of treatment or medical uses of the invention lacks all or a portion of at least of one adenovirus sequences, the cells containing the SC-Ad suitable for use in the methods of treatment or medical uses of the invention can provide the missing adenovirus polypeptide. For example, when the SC-Ad suitable for use in the methods of treatment or medical uses of the invention is designed to lack nucleic acid encoding the adenovirus fiber polypeptide, an adenovirus fiber polypeptide-expressing cell line can be used to generate the SC-Ad such that the SC-Ad suitable foruse in the methods oftreatmentormedicaluses oftheinventioncontains the fiber polypeptide (e.g., the wild-type fiber polypeptide) while lacking the nucleic acid that encodes the fiber polypeptide (e.g., the wild-type fiber polypeptide). For example, when the SC-Ad suitable for use in the methods of treatment or medical uses of the invention is designed to lack nucleic acid encoding the V polypeptide, an adenovirus V polypeptide-expressing cell line can be used to generate the SC-Ad suitable for use in the methods of treatment or medical uses of the invention such that the SC-Ad suitable for use in the methods of treatment or medical uses of the invention contains the V polypeptide (e.g., the wild-type V polypeptide) while lacking the nucleic acid that encodes the V polypeptide (e.g., the wild-type V polypeptide). For example, when the SC-Ad suitable for use in the methods of treatment or medical uses of the invention is designed to lack nucleic acid encoding the pllla polypeptide, an adenovirus pllla polypeptide-expressing cell line can be used to generate the SC-Ad such that the SC-Ad suitable for use in the methods of treatment or medical uses of the invention contains the pllla polypeptide (e.g., the wild-type pllla polypeptide) while lacking the nucleic acid that encodes the pllla polypeptide (e.g., the wild-type pllla polypeptide). In some cases, cells containing SC-Ads suitable for use in the methods of treatment or medical uses of the invention can increase the available number of copies of that virus by at least 100-fold (e.g., by 100-fold to 15,000-fold, by 500- to 10,000-fold, by 5,000- to 10,000-fold, or by 5,000- to 15,000-fold). Avirus can be expandeduntil a desired concentrationis obtained in standard cell culture media (e.g., DMEM or RPMI-1640 supplemented with 5-10% fetal bovine serum at 37°C in 5% CO2). A viral titre typically is assayed by inoculating cells (e.g., A549 or 293 cells) in culture or by quantitating viral genomes by optical density or real-time PCR. In some cases, cells containing a SC-Ad suitable for use in the methods of treatment or medical uses of the invention can be used to propagate the SC-Ad (e.g., to establish a stock of theSC-Ad). For example, a stock of the SC-Ad suitable for use in the methods of treatment or medical uses of the invention can be produced by growth in mammalian cells. In some cases, a stock of the SC-Ad can be aliquoted and frozen, and can be stored at-70°Cto -80°C (e.g., at concentrations higher than the therapeutically effective dose). In some cases, a stock of the SC-Ad suitable foruse in the methods oftreatment or medical uses ofthe invention canbe stored in a stabilizing solution. Examples of stabilizing solutions include, without limitation, sugars (e.g., trehalose, dextrose, and glucose), amino acids, glycerol, gelatin, monosodium glutamate, Ca2+, and Mg2+.

[0262] The invention will be further described in the following examples, which do not limitthe scope of the invention described in the claims. EXAMPLES

[0263] Example 1: A Phase 1, First in Human, Open label, Single Ascending Dose and Multidose Study to Assess the Safety, Reactogenicity, and Immunogenicity of the Adenovirus Vector SARS-CoV-2 Investigational Product SC-Ad6-l (Example of SC-Ad suitable for use in the methods of treatment or medical uses of the invention) given via Intramuscular, Intranasal or Inhaled Administration in Healthy Volunteers (ClinicalTrials.gov Identifier: NCT04839042)

[0264] In a recent three-part, phase 1, first in human, Open label, Single Ascending Dose and Multidose Study SC-Ad6-l was administered to humans via IM, IN or IH administration in healthy volunteers. The main purpose of Part A of this study was examine how safe and effective a single ascending dose (SAD) of SC-Ad6-l was on participants. The purpose of Part B was to examine how safe and effective a multiple dose (MD) of SC-Ad6-l was on participants. Finally, the purpose of Part C was to examine how safe and effective a single booster dose (BD) of SC-Ad6-l was on participants.

[0265] SC-Ad6-l is a single-cycle, adenovirus serotype 6 (SC-Ad6) vector carrying the codon- optimised SARS-CoV-2 spike transgene which encodes for the SARS-CoV-2 spike protein. The pIIIA and E3 genes have been deleted from the parental adenovirus (human adenovirus group C, serotype 6; strain Tonsil 99; American Type Culture Collection [ATCC] VR-1083). The pIIIA gene codes for the pIIIA capsid cement protein that is required for DNA packaging and virus capsid assembly, consequently, its deletion renders the virus unable to form infectious particles in the initially infected host cells. TheE3 gene codes for immune evasion proteins and is also responsible for efficient cell lysis and release of virus progeny, consequently its deletion suppresses viral immune evasion and release of viral progeny.

[0266] Part C relates to will evaluate three SBD dose levels administered IH in a total of 3 cohorts. Unlike for either Part A or Part B, all subjects in Part C will already have been fully vaccinated or fully vaccinated and boosted against COVID-19. Three cohorts (Cohort 12, Cohort 13 and Cohort 14) will receive a single administration of SC-Ad6-l IH (Table 1). Part C will evaluate three dose levels administered via an IH route (Cohorts 12, 13 and 14). The total study duration for each individual participant in Part C of this study is approximately 20 weeks (134 days) which includes:

[0267] • A screening period of up to 4 weeks (28 days)

[0268] • A period of approximately 6 weeks (43 days) commencing with dose administration on Day

[0269] 1 followed by study assessments up to Day 43.

[0270] • An assessment free interval of 9 weeks (63 days) until the EoS visit on Day 106.

[0271] Table 1 : SC-Ad6-l-002 Study IN or IH Cohorts

[0272] Key Inclusion Criteria for Part A, B and C

[0273] • Adult males and females, 18 to 60 years of age (inclusive);

[0274] • Body mass index > 18.0 and < 32.0 kg / m2, with a body weight > 50 kg at screening;

[0275] • Must not have smoked more than 5 cigarettes / week in the past 3 months;

[0276] • Medically healthy without clinically significant abnormalities atthe screening visit, and prior to dose administration on Day 1, including: a. Physical examination without any clinically relevant findings. b. Systolic blood pressure in the range of 90 to 140 mmHg (inclusive) and diastolic blood pressure in the range of 50 to 90 mmHg (inclusive) after 5 minutes in supine position. c. Heart rate in the range of 40 to 100 bpm (inclusive) after 5 minutes rest in supine position. d. Body temperature (tympanic), between 35.5°C and 37.7°C (inclusive). e. No clinically significant findings in serum chemistry, haematology, coagulation and urinalysis tests as judged by the PI.

[0277] • Conventional 12-lead electrocardiogram (ECG) recording in triplicate (the mean of triplicate measurements will be used to determine eligibility at the screening visit and to confirm eligibility pre-dose on Day 1) consistent with normal cardiac conduction and function, including: a. Normal sinus rhythm with HR between 40 and 100 bpm (inclusive). b. QTcF < 450 msec for males and <470 msec for females. c. QRS duration of < 120 msec. d. PR interval of in the range of > 120 msec to < 220 msec. e. Electrocardiogram morphology consistent with healthy cardiac ventricular conduction and normal rhythm. f. No family history of short or long QT syndrome, or sudden cardiac death. g. No history of risk factors for torsade de pointes or a known arrythmia.

[0278] • Female participants: a. Must be of non-childbearing potential i.e., documented surgical sterilisation (hysterectomy, bilateral salpingectomy, bilateral oophorectomy at least 6 weeks before screening) or postmenopausal (where postmenopausal is defined as no menses for 12 months without an alternative medical cause and a follicle- stimulating hormone level > 40 IU7L at the screening visit), OR b. If of childbearing potential, the participant:

[0279] • Must not be breastfeeding and must have a negative pregnancy test at the screening visit (blood test) and before the first study drug administration (Day 1, urine test). They must agree not to attempt to become pregnant, and must agree to: i. Use 2 forms of highly effective contraceptive method (Appendix 3) between signing consent, during the study, and at least 90 days after the last dose of study therapy, OR ii. Use at least one highly effective contraceptive method (Appendix 3), plus an additional barrier-method of contraception from signing the consent form until at least 90 days after the last dose of study drug. Acceptable barrier methods include female diaphragm, condom usage for the male partner.

[0280] • Must agree to not donate ova for at least 90 days after the last dose of study therapy. c. Women of childbearing potential with same sex partners (abstinence from penile- vaginal intercourse) are eligible when this is their preferred and usual lifestyle.

[0281] • Male participants, if not surgically sterilised, must be willing not to donate sperm and, if engaging in sexual intercourse with a female partner who could become pregnant, must be willing to use a condom in addition to having the female partner use a highly effective contraceptive method (Appendix 3) from signing the consent form until at least 90 days after the last dose of study drug.

[0282] • For Part C ONLY: must have been fully vaccinated against CO VID- 19 (SARS-CoV-2) and have received the last dose of CO VID vaccine not less than 3 months prior to IP administration on Day 1 for Cohorts 9 and 10 or must have been fully vaccinated against CO VID- 19 (SARS-CoV-2) and have received the last dose of CO VID vaccine not less than 5 months prior to IP administration on Day 1 for Cohort 11 or must have been fully vaccinated against COVID- 19 (SARS-CoV-2) or boosted following full vaccination and have received the last dose of COVID vaccine not less than 12 months prior to IP administration on Day 1 for Cohorts 12, 13, and 14. Proof of CO VID- 19 vaccination must be provided by documentation (e.g., a copy or picture of vaccination record, etc.).

[0283] Key Exclusion Criteria for Parts A, B and C

[0284] • History or presence of significant cardiovascular, pulmonary, hepatic, renal, haematological, gastrointestinal, endocrine, immunologic, dermatologic or neurological disease, including any acute illness or major surgery within the past 3 months determined by the PI to be clinically relevant;

[0285] • History of chronic respiratory disorders including asthma, emphysema, interstitial lung disease, pulmonary hypertension, recurrent pneumonia, COPD, or recent (< 14 days prior to screening) or ongoing respiratory tract infection (NOTE: If in the judgement of the PI a respiratory disorder is transient, the volunteer may be considered eligible and administration of investigational product (IP) may be deferred but deferral should not exceed 28 days from screening);

[0286] • History of thrombosis (e.g., deep vein thrombosis, pulmonary embolism, cerebral venous sinus thrombosis, etc.), heparin-induced thrombocytopenia and thrombosis syndrome (HITTS), any prothrombotic condition (including but not limited to protein c / s deficiency, antiphospholipid syndrome, factor V Leiden, etc.), any coagulation dysregulation disorder, or a history of thrombosis noted in immediate family members;

[0287] • History of any neurological disorders or seizures including Guillain-Barre syndrome, with the exception of febrile seizures during childhood;

[0288] • History of type 1 or type 2 diabetes;

[0289] • Recent (within the last 2 weeks) or current infection that requires systemically absorbed antibiotic, antifungal, antiparasitic or antiviral medications;

[0290] • For Part C ONLY: For Cohorts 9 and 10, a known previous infection of SARS-CoV- 2 or a positive SARS-CoV-2 PCR test occurring within 3 months prior to IP administration on Day 1 ; for Cohort 11 , a known previous infection of SARS-CoV-2 or a positive SARS-CoV-2 PCR test occurring within 5 months prior to IP administration on Day 1; and for Cohorts 12, 13, and 14, a known previous infection of SARS-CoV-2 within 12 months prior to IP administration on Day 1 or a positive SARS-CoV-2 PCR test occurring within 12 months prior to IP administration on Day 1 ;

[0291] • Part C (IH ONLY): For Cohorts 12, 13 and 14, an anti-Spike IgG concentration of 150,000 AU / mL or higher during screening.

[0292] • For Part C ONLY: planning on receiving an approved SARS-CoV-2 vaccine or boost of a SARS-CoV-2 vaccine within three weeks after study drug dosing for Cohorts 9, 10 and 11 and within six weeks after study drug dosing for Cohorts 12, 13 and 14;

[0293] • Any history of malignant disease < 5 years prior to registration (excludes curatively treated basal cell or squamous cell carcinoma of the skin, carcinoma in situ of the cervix, or papillary thyroid cancer); • History of radiation therapy or cytotoxic / cancer chemotherapy < 5 years prior to IP administration on Day 1 ;

[0294] • Received vaccination with another agent within 30 days prior to IP administration on Day 1;

[0295] • History of clinically relevant immunosuppression from, but not limited to, immunodeficiency conditions such as common variable hypogammaglobulinemia;

[0296] • Use of orplansto use systemic immunosuppressive (e.g., corticosteroids, methotrexate, azathioprine, cyclosporine) or immunomodulating medications (e.g., interferon) during the study or within 3 months prior to IP administration on Day 1 ;

[0297] • Liver function test results elevated more than 1.5-fold above the upper limit of normal (ULN) for gamma-glutamyl transferase [GGT], bilirubin (total, conjugated and unconjugated) or alkaline phosphatase (ALP) or elevated 2.0-fold above the upper limit of normal (ULN) for aspartate aminotransferase (AST) or alanine aminotransferase (ALT). Participants with ALP and / or ALT / AST above the limits specified may be included, at the discretion of the PI, if the levels are unaccompanied by clinical signs and are determined to be normal variants;

[0298] • Positive test results for active human immunodeficiency virus (HIV), hepatitis B surface antigen (HBsAg) or hepatitis C virus (HCV) antibodies at the screening visit;

[0299] • Estimated creatinine clearance (CrCl) < 60 mL / min using the Cockcroft-Gault formula or serum creatinine more than 1.5-fold above the ULN;

[0300] • History of substance abuse or alcohol abuse (defined as more than 14 standard drinks per week or regularly consuming more than 4 standard drinks on any one day; where 1 standard drink is 10 g of pure alcohol and is equivalent to 285 mLbeer [4.9% Ale. / Vol.], 100 mL wine [12% Ale. / Vol.], 30 mL spirit [40% Ale. / Vol.]) during < 12 weeks prior to the screening visit;

[0301] • Positive drug or alcohol test results at the screening visit (may be repeated once, if a positive test was recorded in the first instance, at the discretion of the PI);

[0302] • Use of any systemically absorbed prescription or over-the-counter medication (NOTE: standard dose multivitamins or dietary supplements are allowed) within 7 days or 5 half-lives of the medication (whichever is longer) prior to the first study drug administration, except occasional use of paracetamol (up to a maximum of 4 doses per day of 500-mg paracetamol, and no more than 3g per week); • Demonstrated clinically significant (required intervention, e.g., emergency room visit, epinephrine administration) allergic reactions (e.g., food, drug, or atopic reactions, asthmatic episodes) which, in the opinion of the PI, would interfere with the volunteer’s ability to participate in the trial;

[0303] • Known hypersensitivity to vaccines or their components;

[0304] • For Women of Childbearing Potential, a positive serum pregnancy test at the screening visit or a positive urine pregnancy test (with confirmatory serum pregnancy test) conducted prior to dose administration on Day 1;

[0305] • Donation of blood or plasma within 30 days prior to first study drug administration, or loss of whole blood of more than 500 mL within 30 days prior to Day 1, or receipt of blood or blood-derived products (i.e. immunoglobulins) within 3 months of the administration of IP on Day 1 ;

[0306] • Have received another IP within 30 days prior to the administration of SC-Ad6-l on Day 1 (the IP cannot be an unapproved COVID-19 vaccine);

[0307] • Any other condition or prior therapy that in the opinion of the PI would make the volunteer unsuitable for this study, including inability to cooperate fully with the requirements of the study protocol or likelihood of noncompliance with any study requirements;

[0308] • Is an employee or an immediate relative of the PI or the sponsor.

[0309] Objectives

[0310] Assess the safety and tolerability of single ascending doses of SC-Ad6-l when administered via IM, IN or IH in healthy volunteers (in both unvaccinated and previously COVID-19 vaccinated individuals).

[0311] Evaluate the humoral and cellular immune response to single ascending doses of SC-Ad6-l when administered via IM, IN or IH in healthy volunteers (in both unvaccinated and previously CO VID-19 vaccinated individuals).

[0312] Evaluate the humoral and cellular immune response to multiple doses (two total doses, administered 21 days apart) of SC-Ad6-l when administered via IM injection or IN spray in healthy volunteers (in both unvaccinated and previously COVD-19 vaccinated individuals).

[0313] Immunogenicity Assessments

[0314] The induction of a humoral response to SARS-CoV-2 (spike protein) will be assessed at the timepoints specified in the schedule of assessments (see Table 2) by measuring anti- SARS- CoV-2 (spike protein) antibody titres in the upper respiratory tract mucosa (IgA) and in serum (IgA and IgG). Upper respiratory tract mucosa and saliva samples will also be assessed for total IgAto allowfor assessment ofthe ratio of SARS-CoV-2 IgA antibody versus total IgAantibody in mucus samples.

[0315] Table 2: Schedule of assessments

[0316]

[0317] A micro-neutralisation assay was also performed on blood samples collected to measure SARS-

[0318] CoV-2 neutralising antibody titres and Ad-6 neutralising antibody titres (ADA response).

[0319] PBMC was isolated from a blood sample collected for the assessment of SARS-CoV-2 cell mediated responses. Processing of the blood sample for PBMC isolation was performed by laboratory staff at the clinical facility or by appropriately qualified laboratory vendors. Assays were be used to test for elevated level of specific Th2 related inflammatory cytokines and specific T cell subpopulation activation or over-reactivation (whichmay includebutnotlimited to CD4 T, CD8+ T and Treg cells).

[0320] IgG Spike Antibody Response

[0321] The IgG spike antibody response refers to the production of immunoglobulin G (IgG) antibodies specifically targeting the spike protein of a virus, such as SARS-CoV-2, which is responsible for facilitating viral entry into host cells. A heightened IgG spike antibody response during infection is immensely beneficial as it plays a pivotal role in combating the virus and mitigating the severity of illness. These antibodies neutralize the virus by binding to its spike protein, preventing it from infecting host cells and reducing viral replication within the body. By controlling the viral load, IgG antibodies help alleviate symptoms and facilitate a faster recovery.

[0322] Table 3 presents the impact of a single booster dose (BD) of SC-Ad6-l on healthy volunteers within Cohort 12, revealing the measurable IgG Spike Antibody Response induced by the Sc- Ad. The table showcases the titres of specific antibodies generated in response to the spike protein of SARS-CoV-2. Notably, the data illustrates a consistent elevation in values of IgG spike antibodies among all participants who received the booster dose. This increase is discernible at both Day 22 and Day 43 in comparison to the initial screening or day 1 values. The average rise in values of IgG spike antibodies between Day 1 and Day 22 approximates 30%, while between Day 1 and Day 43 , an even more significant immune response is ob served, with an average increase of approximately 60%. These results provide compelling evidence of the efficacy of SC-Ad6-l in eliciting a robust immune response against SARS-CoV-2.

[0323] Fig 1 illustrates the fold increase of IgG spike antibodies for each volunteer over time. It is evident from figure 1 that each volunteer showed a positive fold increase of IgG spike antibodies. One volunteer even showed an over 5-fold increase in IgG spike antibodies. Moreover, these findings find support in Fig 2, which illustrates the collective response of healthy volunteers who received the booster dose. Fig 2 showcases a mean fold increase of 1 .09 at day 43 of the trial, further reinforcing the SC-Ads's efficacy in augmenting the humoral immune response. This comprehensive analysis underscores the potency of administration of SC-Ad6-l via inhalation in bolstering the body's defence against SARS-CoV-2 infection.

[0324] Table 3 : IgG Spike Antibody Response (absorbance units (AU) / ml)

[0325] IgG Receptor Binding Domain (RBD) Antibody Response

[0326] IgG RBD (Receptor Binding Domain) antibody response refers to the production of immunoglobulin G (IgG) antibodies specifically targeted against the receptor binding domain of a virus, such as on the spike protein of SARS-CoV-2. Having IgG antibodies directed against the receptor binding domain (RBD) of SARS-CoV-2 offers crucial benefits in combatting infection. These antibodies play a pivotal role in neutralizing the virus by binding to its RBD, thereby preventing its attachment to host cells and subsequent infection. Table 4 presents the impact of a single booster dose (BD) of SC-Ad6-l on healthy volunteers within Cohort 12, revealing the measurable IgGRBD antibody response inducedby the Sc- Ad. The table showcases the titres of specific antibodies generated in response to the RBD of SARS-CoV-2. Notably, the data shows a consistent elevation in values of IgG spike antibodies among almost all participants who received the booster dose. The average rise in values of IgG spike antibodies between Day 1 and Day 22 and between Day 1 and Day 43 is approximately 25%. Again this highlights, the SC-Ads’ efficacy in augmenting the humoral immune response.

[0327] Fig 3 illustrates the fold increase of IgG RBD antibodies for each volunteer overtime. It is evident from figure 3 that almost all volunteers showed a positive fold increase of IgG RBD antibodies. One volunteer even showed an over 4-fold increase in IgG RBD antibodies. Fig 4 further highlights this increase in IgGRBD antibodies showing a positive correlation in average fold increase of antibodies across all participants over time.

[0328] Table 4: IgG RBD Antibody Response (absorbance units (AU) / ml)

[0329] IgG nucleocapsid Antibody Response

[0330] "IgG nucleocapsid Antibody Response" refers to the production of IgG antibodies specific to the nucleocapsid protein of SARS-CoV-2.

[0331] The SC-Ad utilised in this clinical trial encodes the SARS-CoV-2 spike protein as an immunogen. It is therefore reasonable to expect no significant increase in IgG nucleocapsid antibodies, since there is no generation of a nucleocapsid immunogen associated with the treatment using the SC-Ad. The nucleocapsid data therefore provide an effective negative control, demonstrating that antibody responses to non-boosted immunogens decrease further over time in the absence of a boost.

[0332] Table 5 presents the impact of a single booster dose (BD) of SC-Ad6-l on healthy volunteers within Cohort 12, revealing the measurable IgG nucleocapsid Antibody Response induced by the SC-Ad. The table illustrates that the titres of specific antibodies generated in response to the nucleocapsid of SARS-CoV-2 shows no significant increase in the values but rather shows there was a slight decrease of 14% between Day 1 and Day 22 and an 4% decrease between Day 22 and Day 43. This potentially highlights how in the absence of a vaccination boost relevant to a particular target the subjects antibodies will steadily decline over time. This highlights the need for effective vaccination boosts.

[0333] Fig 5 illustrates the fold increase of IgG nucleocapsid antibodies for each volunteer overtime. It is evident from figure 5 that almost all volunteers showed a negligent or negative correlation of IgG nucleocapsid antibodies over time. Fig 6 further supports this by showing a corresponding minor fold average decrease over time. Table 5: IgG nucleocapsid Antibody Response (absorbance units (AU) / ml)

[0334] Example 2

[0335] Generation of anti-spike protein antibodies in response to treatment regimens using inhalation administration of SC-Ads

[0336] The generation of anti-spike protein antibodies in response to different treatment regiments using inhalation administration of SC-Ads was investigated in three Cohorts (Cohorts 12 and 13, as described above, and Cohort 15). Details of the treatments administered were as set out in Table 6 below.

[0337] Table 6

[0338] Mean levels of anti-spike protein antibodies were assessed in serum (IgG and IgA), upper respiratory tract mucosa (IgA) and saliva samples (IgA). The results obtained are set out in Figs 7 and 8, and discussed below.

[0339] IgG anti-spike protein antibodies in serum samples

[0340] As can be seen in Fig 7, Cohorts 12 and 13 (each receiving a single administration of SC-Ad) both exhibited similar patterns of IgG in the serum overtime. In both cases, the mean total amount of serum IgG increasing from 0 to 43 days. In the case of Cohort 12, the mean amount of serum IgG remained stable from 43 days to 106 days, while in the case of Cohort 13 a slight decrease in the level was observed over this period. Mean serum IgG present was greater in Cohort 12 than in Cohort 13 at all timepoints assessed. In Cohort 12, mean serum IgG peaked at an increase of just over 1-fold as compared to baseline. In Cohort 13, mean serum IgG peaked at an increase of approximately 0.66-fold as compared to baseline at day 43, before dropping to approximately 0.4-fold as compared to basely by day 106. This increased ability of Cohort 12 to stimulate IgG antibody formation in serum may be considered surprising, given that Cohort 12 received a smaller dose of SC-Ads than did Cohort 13.

[0341] Cohort 15 (receiving two administrations of SC-Ad) achieved a higher increase in serum IgG than either Cohort 12 or Cohort 13. In Cohort 15, the mean amount of serum IgG peaked at an increase of approximately 3.1 -fold over baseline at 22 days post-administration. This was then followed by slight decrease at the next timepoint assessed. The levels then returned to approximately peakvalues, gradually reducingto an increase of approximately 2.4-fold over baseline at day 106.

[0342] The results obtained in respect of Cohort 15 illustrate the ability of a treatment regimen using multiple administrations to achieve a long-lasting IgG antibody response in serum.

[0343] IgA anti-spike protein antibodies in serum, nasal and saliva sample

[0344] Serum IgA anti-spike protein antibody levels

[0345] Fig 8 A compares the mean levels of anti-spike protein IgA antibodies observed in serum in Cohorts 12, 13 and 15.

[0346] Both Cohorts 12 and 13 demonstrated a marked increase in IgA antibody levels over the first 22 days, with both achieving an approximately 1-fold increase as compared to baseline at this timepoint. Over time, the amount of IgA anti-spike protein antibodies in samples from Cohort 13 decreased markedly. In contrast, samples from Cohort 12 demonstrated a slight increase in IgA anti-spike protein levels from day 22 to 43, before a dramatic increase between days 43 and 106, reaching approximately 3.75-fold increase as compared to baselin. As with the IgG antibodies reported on above, serum levels of anti-spike protein IgAantibodies in samples from Cohort 12 were higher than those from Cohort 13 at all timepoints tested.

[0347] Again, these results indicate thatthe lower dose of SC-Ad used in Cohort 12 gave rise to longer- lasting serum antibody protection than did Cohort 13. Such doses may provide a favoured approach in clinical contexts in which it is wished to develop a strong systemic immune response by means of a single administration of SC-Ads. Turning to Cohort 15, the results in Fig 8 A illustrate that an immediate response was observed, with peak concentration of anti-spike protein IgA antibodies achieved in the first 23 to 30 days. The highest antibody levels obtained were increased approximately 5-fold as compared to baseline. The level of antibodies then decreasedup to 43 days, before remainingbroadly stable until day 106.

[0348] Serum levels of IgAanti-spike protein antibodies achievedat day 106 were broadly comparable with those observed in respect of Cohort 12. Accordingly, a multi-dose approach as in Cohort 15 may provide a suitable alternative strategy in the case that is wished to develop a strong systemic immune response by means of a plurality of administrations of SC-Ads.

[0349] Nasal IgA anti-spike protein antibody levels

[0350] Fig 8B compares the mean levels of anti-spike protein IgA antibodies observed in nasal mucosal samples in Cohorts 12, 13 and 15.

[0351] Cohorts 13 and 15 both exhibit a sharp increase in antibody levels over the first 22 days, resulting in respectively a 1-fold and 1.5-fold increase as compared to baseline. The levels observed in respect of Cohort 15 then increase slightly, before decreasing and then returning to an approximately 1-fold increase.

[0352] Cohort 12 demonstrates a lower level of antibodies at day 22 (increased approximately 0.3- fold as compared to baseline), which then decreases at day 43. However, from days 43 to 106 the level of antibodies markedly increases, with levels of IgA anti-spike protein antibodies observed at this timepoint being more than 5-fold increased as compared to baseline.

[0353] Saliva sample IgA anti-spike protein antibody levels

[0354] Fig 8B compares the mean levels of anti-spike protein IgA antibodies observed in nasal mucosal samples in Cohorts 12, 13 and 15.

[0355] Cohorts 12 and 15 respectively exhibit a 1-fold and 1.5-fold increase in antibody levels, as compared to baseline, over the first 22 days. Both then demonstrate a reduction in levels, with Cohort 15 decreasing to an approximately 0.5-fold increase as compared to baseline, before rising to an increase of 0.7-fold by day 106. In contrast, while Cohort 12 decreases to approximately 0.8-fold increased as comparedto baseline by day 43, this then rises to a more than 2-fold increase by day 106. Cohort l3 exhibits lower values than either Cohort 12 or 15 at days 22 and 43 (approximately 0.5-fold and 0.25-fold increased versus baseline respectively), before levels rise to a 2-fold increase by day 106.

[0356] Taken together, these data suggest that treatment regimens utilising a single dose of SC-Ads may be highly effective in achieving an IgA antibody response in saliva.

[0357] SEQUENCE INFORMATION

[0358] The following sequences are referred to in the present specification.

[0359] SEQ ID NO:1 - amino acid sequence of a SARS-CoV-2 Spike polypeptide

[0360] MFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFF SNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSL

[0361] LIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQP FLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGI NITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVD CALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASV YAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEV RQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPF ERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPA TVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDP QTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYST GSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRARSVASQSIIAYTM SLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYG

[0362] SFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSKPSKRS FIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYT SALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAI GKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDKVEA EVQIDRLITGRLQ SLQTYVTQQLIRAAEIRASANLAATKMSEC VLGQ SKRVDFCGKG YHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWF VTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTS PDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWYIWLG FIAGLIAIVMVTIMLCCMTSCCSCLKGCCSCGSCCKFDEDDSEPVLKGVKLHYT

[0363] SEQ ID NO:2 amino acid sequence of a SARS-CoV-2 Spike polypeptide lacking an ER retention sequence

[0364] MFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFF SNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSL LIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQP FLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGI NITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVD CALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASV YAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEV RQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPF ERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPA TVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDP QTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYST

[0365] GSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRARSVASQSIIAYTM SLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYG SFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSKPSKRS FIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYT SALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAI GKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDKVEA EVQIDRLITGRLQ SLQTYVTQQLIRAAEIRASANLAATKMSEC VLGQ SKRVDFCGKG YHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWF VTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTS PDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWYIWLG FIAGLIAIVMVTIMLCCMTSCCSCLKGCCSCGSCCKFDEDDSEPVLKGV

[0366] SEQ ID NO:3 amino acid sequence of an ectodomain of a SARS-CoV-2 Spike polypeptide

[0367] MFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFF SNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSL

[0368] LIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQP FLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGI NITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVD CALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASV YAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEV RQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPF ERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPA TVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDP QTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYST GSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRARSVASQSIIAYTM

[0369] SLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYG SFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSKPSKRS FIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYT SALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAI GKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDKVEA EVQIDRLITGRLQ SLQTYVTQQLIRAAEIRASANLAATKMSEC VLGQ SKRVDFCGKG YHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWF VTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTS PDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYI

[0370] SEQ ID NO:4 amino acid sequence of a receptor binding domain of a SARS-CoV-2 Spike polypeptide

[0371] MFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFF SNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSL

[0372] LIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQP FLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGI NITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVD CALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPF

[0373] SEQ ID NO:5 amino acid sequence of a receptor binding domain of a SARS-CoV-2 Spike polypeptide fused to an Ig polypeptide

[0374] DPLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESI VRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSP TKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNL

[0375] DSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQP TNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLCCVECPPCPAPPVAGPSVFLFPPK PKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRV VSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSR

[0376] WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0377] SEQ ID NO:6 amino acid sequence of a receptor binding domain of a SARS-CoV-2 Spike polypeptide fused to a streptavidin polypeptide

[0378] DPLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESI VRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSP TKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNL DSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQP TNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLCCVECPPCPAPPVAGTMGGAAGS GAAEAGITGTWYNQLGSTFIVTAGADGALTGTYESAVGNAESRYVLTGRYESAPATD

[0379] GSGTALGWTVAWKNNYRNAHSATTWSGQYVGGAEARINTQWLLTSGTTEANAWKS TLVGHDTFTKVKPSAASGS

[0380] SEQ ID NO:7 amino acid sequence of a receptor binding domain of a SARS-CoV-2 Spike polypeptide fused to a sigma coil polypeptide

[0381] DPLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESI VRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSP TKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNL DSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQP TNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLCCVECPPCPAPPVAGSKGLESRVS ALEKTSQIHSDTILRITQGLDDANKRIIALEQSRDDLVASVSDAQLAISRLESSIGALQT

[0382] VVNGLDSSVTQLGARVGQLETGLAELRVDHDNLVARVDTAERNIGSLTTELSTLTLRV TSI

[0383] SEQ ID NO:8 amino acid sequence of an ectodomain of an ACE2 chaff polypeptide

[0384] MSSSSWLLLSLVAVTAAQSTIEEQAKTFLDKFNHEAEDLFYQSSLASWNYNTNITEEN VQNMNNAGDKWSAFLKEQSTLAQMYPLQEIQNLTVKLQLQALQQNGSSVLSEDKS KRLNTILNTMSTIYSTGKVCNPDNPQECLLLEPGLNEIMANSLDYNERLWAWESWRS EVGKQLRPLYEEYVVLKNEMARANHYEDYGDYWRGDYEVNGVDGYDYSRGQLIE DVEHTFEEIKPLYEHLHAYVRAKLMNAYPSYISPIGCLPAHLLGDMWGRFWTNLYSLT VPFGQKPNIDVTDAMVDQAWDAQRIFKEAEKFFVSVGLPNMTQGFWENSMLTDPG

[0385] NVQKAVCHPTAWDLGKGDFRILMCTKVTMDDFLTAHHEMGHIQYDMAYAAQPFLL RNGANEGFHEAVGEIMSLSAATPKHLKSIGLLSPDFQEDNETEINFLLKQALTIVGTLP FTYMLEKWRWMVFKGEIPKDQWMKKWWEMKREIVGVVEPVPHDETYCDPASLFH VSNDYSFIRYYTRTLYQFQFQEALCQAAKHEGPLHKCDISNSTEAGQKLFNMLRLGK SEPWTLALENVVGAKNMNVRPLLNYFEPLFTWLKDQNKNSFVGWSTDWSPYADQS IKVRISLKSALGDKAYEWNDNEMYLFRSSVAYAMRQYFLKVKNQMILFGEEDVRVA

[0386] NLKPRISFNFFVTAPKNVSDIIPRTEVEKAIRMSRSRINDAFRLNDNSLEFLGIQPTLGPP NQPPVS

[0387] SEQ ID NO:9 amino acid sequence of an inactivated ectodomain of an ACE2 chaff polypeptide

[0388] MSSSSWLLLSLVAVTAAQSTIEEQAKTFLDKFNHEAEDLFYQSSLASWNYNTNITEEN VQNMNNAGDKWSAFLKEQSTLAQMYPLQEIQNLTVKLQLQALQQNGSSVLSEDKS KRLNTILNTMSTIYSTGKVCNPDNPQECLLLEPGLNEIMANSLDYNERLWAWESWRS EVGKQLRPLYEEYVVLKNEMARANHYEDYGDYWRGDYEVNGVDGYDYSRGQLIE DVEHTFEEIKPLYEHLHAYVRAKLMNAYPSYISPIGCLPAHLLGDMWGRFWTNLYSLT VPFGQKPNIDVTDAMVDQAWDAQRIFKEAEKFFVSVGLPNMTQGFWENSMLTDPG

[0389] NVQKAVCLPTAWDLGKGDFRILMCTKVTMDDFLTAHHEMGHIQYDMAYAAQPFLL RNGANEGFHEAVGEIMSLSAATPKHLKSIGLLSPDFQEDNETEINFLLKQALTIVGTLP FTYMLEKWRWMVFKGEIPKDQWMKKWWEMKREIVGVVEPVPHDETYCDPASLFH VSNDYSFIRYYTRTLYQFQFQEALCQAAKHEGPLHKCDISNSTEAGQKLFNMLRLGK SEPWTLALENVVGAKNMNVRPLLNYFEPLFTWLKDQNKNSFVGWSTDWSPYADQS IKVRISLKSALGDKAYEWNDNEMYLFRSSVAYAMRQYFLKVKNQMILFGEEDVRVA NLKPRISFNFFVTAPKNVSDIIPRTEVEKAIRMSRSRINDAFRLNDNSLEFLGIQPTLGPP NQPPVS

[0390] SEQ ID NO:10 amino acid sequence of a TcdA / B fusion polypeptide

[0391] MPSSVSWGILLLAGLCCLVPVSLAEDPMPSSVSWGILLLAGLCCLVPVSLAEDPFNLV TGWQTINGKKYYFDINTGAALISYKIINGKHFYFNNDGVMQLGVFKGPDGFEYFAPA NTQNNNIEGQAIVYQSKFLTLNGKKYYFDQDSKAVTGWRIINNEKYYFNPNNAIAAV GLQVIDNNKYYFNPDTAIISKGWQTVQGSRYYFDTDTAIAFNGYKTIDGKHFYFDSD CVVKIGVFSTSNGFEYFAPANTYNNNIEGQAIVYQSKFLT

[0392] SEQ ID NO:11 amino acid sequence of a SARS-CoV-2 ORFlab polypeptide

[0393] MESLVPGFNEKTHVQLSLPVLQVRDVLVRGFGDSVEEVLSEARQHLKDGTCGLVEV EKGVLPQLEQPYVFIKRSDARTAPHGHVMVELVAELEGIQYGRSGETLGVLVPHVGEI PVAYRKVLLRKNGNKGAGGHSYGADLKSFDLGDELGTDPYEDFQENWNTKHSSGV TRELMRELNGGAYTRYVDNNFCGPDGYPLECIKDLLARAGKASCTLSEQLDFIDTKR GVYCCREHEHEIAWYTERSEKSYELQTPFEIKLAKKFDTFNGECPNFVFPLNSIIKTIQP RVEKKKLDGFMGRIRSVYPVASPNECNQMCLSTLMKCDHCGETSWQTGDFVKATCE FCGTENLTKEGATTCGYLPQNAVVKIYCPACHNSEVGPEHSLAEYHNESGLKTILRKG GRTIAFGGCVFSYVGCHNKCAYWVPRASANIGCNHTGVVGEGSEGLNDNLLEILQK EKVNINIVGDFKLNEEIAIILASFSASTSAFVETVKGLDYKAFKQIVESCGNFKVTKGK AKKGAWNIGEQKSILSPLYAFASEAARVVRSIFSRTLETAQNSVRVLQKAAITILDGISQ YSLRLIDAMMFTSDLATNNLVVMAYITGGVVQLTSQWLTNIFGTVYEKLKPVLDWLE EKFKEGVEFLRDGWEIVKFISTCACEIVGGQIVTCAKEIKESVQTFFKLVNKFLALCA DSIIIGGAKLKALNLGETFVTHSKGLYRKCVKSREETGLLMPLKAPKEIIFLEGETLPT EVLTEEVVLKTGDLQPLEQPTSEAVEAPLVGTPVCINGLMLLEIKDTEKYCALAPNM MVTNNTFTLKGGAPTKVTFGDDTVIEVQGYKSVNITFELDERIDKVLNEKCSAYTVE LGTEVNEFAC WAD AVIK TLQPVSELLTPLGIDLDEWSMATYYLFDESGEFKLASHMY CSFYPPDEDEEEGDCEEEEFEPSTQYEYGTEDDYQGKPLEFGATSAALQPEEEQEED WLDDDSQQTVGQQDGSEDNQTTTIQTIVEVQPQLEMELTPVVQTIEVNSFSGYLKLT DNVYIKNADIVEEAKKVKPTVVVNAANVYLKHGGGVAGALNKATNNAMQVESDD YIATNGPLKVGGSCVLSGHNLAKHCLHVVGPNVNKGEDIQLLKSAYENFNQHEVLL APLLSAGIFGADPIHSLRVCVDTVRTNVYLAVFDKNLYDKLVSSFLEMKSEKQVEQKI AEIPKEEVKPFITESKPSVEQRKQDDKKIKACVEEVTTTLEETKFLTENLLLYIDINGNL HPDSATLVSDIDITFLKKDAPYIVGDVVQEGVLTAVVIPTKKAGGTTEMLAKALRKVP TDNYITTYPGQGLNGYTVEEAKTVLKKCKSAFYILPSIISNEKQEILGTVSWNLREML AHAEETRKLMPVCVETKAIVSTIQRKYKGIKIQEGVVDYGARFYFYTSKTTVASLINT LNDLNETLVTMPLGYVTHGLNLEEAARYMRSLKVPATVSVSSPDAVTAYNGYLTSSS KTPEEHFIETISLAGSYKDWSYSGQSTQLGIEFLKRGDKSVYYTSNPTTFHLDGEVITF DNLKTLLSLREVRTIKVFTTVDNINLHTQVVDMSMTYGQQFGPTYLDGADVTKIKPH NSHEGKTFYVLPNDDTLRVEAFEYYHTTDPSFLGRYMSALNHTKKWKYPQVNGLTSI KWADNNCYLATALLTLQQIELKFNPPALQDAYYRARAGEAANFCALILAYCNKTVGE LGDVRETMSYLFQHANLDSCKRVLNVVCKTCGQQQTTLKGVEAVMYMGTLSYEQF KKGVQIPCTCGKQATKYLVQQESPFVMMSAPPAQYELKHGTFTCASEYTGNYQCGH

[0394] YKHITSKETLYCIDGALLTKSSEYKGPITDVFYKENSYTTTIKPVTYKLDGVVCTEIDP KLDNYYKKDNSYFTEQPIDLVPNQPYPNASFDNFKFVCDNIKFADDLNQLTGYKKPA SRELKVTFFPDLNGDVVAIDYKHYTPSFKKGAKLLHKPIVWHVNNATNKATYKPNT

[0395] WCIRCLWSTKPVETSNSFDVLKSEDAQGMDNLACEDLKPVSEEVVENPTIQKDVLEC

[0396] NVKTTEVVGDIILKPANNSLKITEEVGHTDLMAAYVDNSSLTIKKPNELSRVLGLKTL

[0397] ATHGLAAVNSVPWDTIANYAKPFLNKVVSTTTNIVTRCLNRVCTNYMPYFFTLLLQL CTFTRSTNSRIKASMPTTIAKNTVKSVGKFCLEASFNYLKSPNFSKLINIIIWFLLLSVC

[0398] LGSLIYSTAALGVLMSNLGMPSYCTGYREGYLNSTNVTIATYCTGSIPCSVCLSGLDS

[0399] LDTYPSLETIQITISSFKWDLTAFGLVAEWFLAYILFTRFFYVLGLAAIMQLFFSYFAVH

[0400] FISNSWLMWLIINLVQMAPISAMVRMYIFFASFYYVWKSYVHVVDGCNSSTCMMCY

[0401] KRNRATRVECTTIVNGVRRSFYVYANGGKGFCKLHNWNCVNCDTFCAGSTFISDEVA

[0402] RDLSLQFKRPINPTDQSSYIVDSVTVKNGSIHLYFDKAGQKTYERHSLSHFVNLDNLR

[0403] ANNTKGSLPINVIVFDGKSKCEESSAKSASVYYSQLMCQPILLLDQALVSDVGDSAEV

[0404] AVKMFDAYVNTFSSTFNVPMEKLKTLVATAEAELAKNVSLDNVLSTFISAARQGFVD

[0405] SDVETKDVVECLKLSHQSDIEVTGDSCNNYMLTYNKVENMTPRDLGACIDCSARHIN

[0406] AQVAKSHNIALIWNVKDFMSLSEQLRKQIRSAAKKNNLPFKLTCATTRQVVNVVTTK

[0407] IALKGGKIVNNWLKQLIKVTLVFLFVAAIFYLITPVHVMSKHTDFSSEIIGYKAIDGGV

[0408] TRDIASTDTCFANKHADFDTWFSQRGGSYTNDKACPLIAAVITREVGFVVPGLPGTIL

[0409] RTTNGDFLHFLPRVFSAVGNICYTPSKLIEYTDFATSACVLAAECTIFKDASGKPVPYC

[0410] YDTNVLEGSVAYESLRPDTRYVLMDGSIIQFPNTYLEGSVRVVTTFDSEYCRHGTCER

[0411] SEAGVCVSTSGRWVLNNDYYRSLPGVFCGVDAVNLLTNMFTPLIQPIGALDISASIVA

[0412] GGIVAIVVTCLAYYFMRFRRAFGEYSHVVAFNTLLFLMSFTVLCLTPVYSFLPGVYSVI

[0413] YLYLTFYLTNDVSFLAHIQWMVMFTPLVPFWITIAYIICISTKHFYWFFSNYLKRRVVF

[0414] NGVSFSTFEEAALCTFLLNKEMYLKLRSDVLLPLTQYNRYLALYNKYKYFSGAMDTT

[0415] SYREAACCHLAKALNDFSNSGSDVLYQPPQTSITSAVLQSGFRKMAFPSGKVEGCMV

[0416] QVTCGTTTLNGLWLDDVVYCPRHVICTSEDMLNPNYEDLLIRKSNHNFLVQAGNVQ

[0417] LRVIGHSMQNCVLKLKVDTANPKTPKYKFVRIQPGQTFSVLACYNGSPSGVYQCAM

[0418] RPNFTIKGSFLNGSCGSVGFNIDYDCVSFCYMHHMELPTGVHAGTDLEGNFYGPFVD

[0419] RQTAQAAGTDTTITVNVLAWLYAAVINGDRWFLNRFTTTLNDFNLVAMKYNYEPLTQ

[0420] DHVDILGPLSAQTGIAVLDMCASLKELLQNGMNGRTILGSALLEDEFTPFDVVRQCS

[0421] GVTFQSAVKRTIKGTHHWLLLTILTSLLVLVQSTQWSLFFFLYENAFLPFAMGIIAMSA

[0422] FAMMFVKHKHAFLCLFLLPSLATVAYFNMVYMPASWVMRIMTWLDMVDTSLSGFK

[0423] LKDCVMYASAVVLLILMTARTVYDDGARRVWTLMNVLTLVYKVYYGNALDQAISM

[0424] WALIISVTSNYSGVVTTVMFLARGIVFMCVEYCPIFFITGNTLQCIMLVYCFLGYFCTC

[0425] YFGLFCLLNRYFRLTLGVYDYLVSTQEFRYMNSQGLLPPKNSIDAFKLNIKLLGVGGK

[0426] PCIKVATVQSKMSDVKCTSVVLLSVLQQLRVESSSKLWAQCVQLHNDILLAKDTTEA

[0427] FEKMVSLLSVLLSMQGAVDINKLCEEMLDNRATLQAIASEFSSLPSYAAFATAQEAYE

[0428] QAVANGDSEVVLKKLKKSLNVAKSEFDRDAAMQRKLEKMADQAMTQMYKQARSE

[0429] DKRAKVTSAMQTMLFTMLRKLDNDALNNIINNARDGCVPLNIIPLTTAAKLMVVIPD

[0430] YNTYKNTCDGTTFTYASALWEIQQVVDADSKIVQLSEISMDNSPNLAWPLIVTALRA

[0431] NSAVKLQNNELSPVALRQMSCAAGTTQTACTDDNALAYYNTTKGGRFVLALLSDLQ

[0432] DLKWARFPKSDGTGTIYTELEPPCRFVTDTPKGPKVKYLYFIKGLNNLNRGMVLGSL

[0433] AATVRLQAGNATEVPANSTVLSFCAFAVDAAKAYKDYLASGGQPITNCVKMLCTHT

[0434] GTGQAITVTPEANMDQESFGGASCCLYCRCHIDHPNPKGFCDLKGKYVQIPTTCAND

[0435] PVGFTLKNTVCTVCGMWKGYGCSCDQLREPMLQSADAQSFLNRVCGVSAARLTPC

[0436] GTGTSTDVVYRAFDIYNDKVAGFAKFLKTNCCRFQEKDEDDNLIDSYFVVKRHTFSN

[0437] YQHEETIYNLLKDCPAVAKHDFFKFRIDGDMVPHISRQRLTKYTMADLVYALRHFDE

[0438] GNCDTLKEILVTYNCCDDDYFNKKDWYDFVENPDILRVYANLGERVRQALLKTVQF

[0439] CDAMRNAGIVGVLTLDNQDLNGNWYDFGDFIQTTPGSGVPVVDSYYSLLMPILTLTR

[0440] ALTAESHVDTDLTKPYIKWDLLKYDFTEERLKLFDRYFKYWDQTYHPNCVNCLDDR CILHCANFNVLFSTVFPPTSFGPLVRKIFVDGVPFVVSTGYHFRELGVVHNQDVNLHS

[0441] SRLSFKELLVYAADPAMHAASGNLLLDKRTTCFSVAALTNNVAFQTVKPGNFNKDFY DFAVSKGFFKEGSSVELKHFFFAQDGNAAISDYDYYRYNLPTMCDIRQLLFVVEVVD KYFDCYDGGCINANQVIVNNLDKSAGFPFNKWGKARLYYDSMSYEDQDALFAYTK RNVIPTITQMNLKYAISAKNRARTVAGVSICSTMTNRQFHQKLLKSIAATRGATVVIGT SKFYGGWHNMLKTVYSDVENPHLMGWDYPKCDRAMPNMLRIMASLVLARKHTTC CSLSHRFYRLANECAQVLSEMVMCGGSLYVKPGGTSSGDATTAYANSVFNICQAVTA NVNALLSTDGNKIADKYVRNLQHRLYECLYRNRDVDTDFVNEFYAYLRKHFSMMIL SDDAVVCFNSTYASQGLVASIKNFKSVLYYQNNVFMSEAKCWTETDLTKGPHEFCSQ HTMLVKQGDDYVYLPYPDPSRILGAGCFVDDIVKTDGTLMIERFVSLAIDAYPLTKHP NQEYADVFHLYLQYIRKLHDELTGHMLDMYSVMLTNDNTSRYWEPEFYEAMYTPHT VLQAVGACVLCNSQTSLRCGACIRRPFLCCKCCYDHVISTSHKLVLSVNPYVCNAPG CDVTDVTQLYLGGMSYYCKSHKPPISFPLCANGQVFGLYKNTCVGSDNVTDFNAIAT CDWTNAGDYILANTCTERLKLFAAETLKATEETFKLSYGIATVREVLSDRELHLSWE VGKPRPPLNRNYVFTGYRVTKNSKVQIGEYTFEKGDYGDAVVYRGTTTYKLNVGDY FVLTSHTVMPLSAPTLVPQEHYVRITGLYPTLNISDEFSSNVANYQKVGMQKYSTLQG PPGTGKSHFAIGLALYYPSARIVYTACSHAAVDALCEKALKYLPIDKCSRIIPARARVE CFDKFKVNSTLEQYVFCTVNALPETTADIVVFDEISMATNYDLSVVNARLRAKHYVY IGDPAQLPAPRTLLTKGTLEPEYFNSVCRLMKTIGPDMFLGTCRRCPAEIVDTVSALVY DNKLKAHKDKSAQCFKMFYKGVITHDVSSAINRPQIGVVREFLTRNPAWRKAVFISP YNSQNAVASKILGLPTQTVDSSQGSEYDYVIFTQTTETAHSCNVNRFNVAITRAKVGIL CIMSDRDLYDKLQFTSLEIPRRNVATLQAENVTGLFKDCSKVITGLHPTQAPTHLSVD TKFKTEGLCVDIPGIPKDMTYRRLISMMGFKMNYQVNGYPNMFITREEAIRHVRAWI GFDVEGCHATREAVGTNLPLQLGFSTGVNLVAVPTGYVDTPNNTDFSRVSAKPPPGD QFKHLIPLMYKGLPWNVVRIKIVQMLSDTLKNLSDRVVFVLWAHGFELTSMKYFVKI GPERTCCLCDRRATCFSTASDTYACWHHSIGFDYVYNPFMIDVQQWGFTGNLQSNH DLYCQVHGNAHVASCDAIMTRCLAVHECFVKRVDWTIEYPIIGDELKINAACRKVQH MVVKAALLADKFPVLHDIGNPKAIKCVPQADVEWKFYDAQPCSDKAYKIEELFYSY ATHSDKFTDGVCLFWNCNVDRYPANSIVCRFDTRVLSNLNLPGCDGGSLYVNKHAF HTPAFDKSAFVNLKQLPFFYYSDSPCESHGKQVVSDIDYVPLKSATCITRCNLGGAVC RHHANEYRLYLDAYNMMISAGFSLWVYKQFDTYNLWNTFTRLQSLENVAFNVVNK GHFDGQQGEVPVSIINNTVYTKVDGVDVELFENKTTLPVNVAFELWAKRNIKPVPEV KILNNLGVDIAANTVIWDYKRDAPAHISTIGVCSMTDIAKKPTETICAPLTVFFDGRVD GQVDLFRNARNGVLITEGSVKGLQPSVGPKQASLNGVTLIGEAVKTQFNYYKKVDG VVQQLPETYFTQSRNLQEFKPRSQMEIDFLELAMDEFIERYKLEGYAFEHIVYGDFSH SQLGGLHLLIGLAKRFKESPFELEDFIPMD STVKNYFITD AQTGSSKC VC SVIDLLLDD FVEIIKSQDLSVVSKVVKVTIDYTEISFMLWCKDGHVETFYPKLQSSQAWQPGVAMP NLYKMQRMLLEKCDLQNYGDSATLPKGIMMNVAKYTQLCQYLNTLTLAVPYNMRVI HFGAGSDKGVAPGTAVLRQ WLPTGTLLVD SDLNDF VSD AD STLIGDC ATVHTANKW DLIISDMYDPKTKNVTKENDSKEGFFTYICGFIQQKLALGGSVAIKITEHSWNADLYK LMGHFAWWTAFVTNVNASSSEAFLIGCNYLGKPREQIDGYVMHANYIFWRNTNPIQ LSSYSLFDMSKFPLKLRGTAVMSLKEGQINDMILSLLSKGRLIIRENNRVVISSDVLVN N

[0442] SEQ ID NO:12 amino acid sequence of a SARS-CoV-2 S polypeptide

[0443] MFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFF SNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSL LIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQP FLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGI NITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVD CALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASV YAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEV RQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPF

[0444] ERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPA

[0445] TVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDP

[0446] QTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYST

[0447] GSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRARSVASQSIIAYTM

[0448] SLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYG

[0449] SFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSKPSKRS

[0450] FIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYT

[0451] SALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAI

[0452] GKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDKVEA

[0453] EVQIDRLITGRLQ SLQTYVTQQLIRAAEIRASANLAATKMSEC VLGQ SKRVDFCGKG

[0454] YHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWF

[0455] VTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTS

[0456] PDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWYIWLG

[0457] FIAGLIAIVMVTIMLCCMTSCCSCLKGCCSCGSCCKFDEDDSE

[0458] SEQ ID NO:13 amino acid sequence of a SARS-CoV-2 ORF3 polypeptide

[0459] MDLFMRIFTIGTVTLKQGEIKDATPSDFVRATATIPIQASLPFGWLIVGVALLAVFQSAS

[0460] KIITLKKRWQLALSKGVHFVCNLLLLFVTVYSHLLLVAAGLEAPFLYLYALVYFLQSIN

[0461] FVRIIMRLWLCWKCRSKNPLLYDANYFLCWHTNCYDYCIPYNSVTSSIVITSGDGTTS

[0462] PISEHDYQIGGYTEKWESGVKDCVVLHSYFTSDYYQLYSTQLSTDTGVEHVTFFIYN

[0463] KIVDEPEEHVQIHTIDGSSGVVNPVMEPIYDEPTTTTSVPL

[0464] SEQ ID NO:14 amino acid sequence of a SARS-CoV-2 E polypeptide

[0465] MYSFVSEETGTLIVNSVLLFLAFVVFLLVTLAILTALRLCAYCCNIVNVSLVKPSFYVY SRVKNLNSSRVPDLLV

[0466] SEQ ID NO:15 amino acid sequence of a SARS-CoV-2 M polypeptide

[0467] MADSNGTITVEELKKLLEQWNLVIGFLFLTWICLLQFAYANRNRFLYIIKLIFLWLLWP

[0468] VTLACFVLAAVYRINWITGGIAIAMACLVGLMWLSYFIASFRLFARTRSMWSFNPETN

[0469] ILLNVPLHGTILTRPLLESELVIGAVILRGHLRIAGHHLGRCDIKDLPKEITVATSRTLSY

[0470] YKLGASQRVAGDSGFAAYSRYRIGNYKLNTDHSSSSDNIA

[0471] SEQ ID NO:16 amino acid sequence of a SARS-CoV-2 ORF3 polypeptide

[0472] MFHLVDFQVTIAEILLIIMRTFKVSIWNLDYIINLIIKNLSKSL

[0473] SEQ ID NO:17 amino acid sequence of a SARS-CoV-2 ORF7 polypeptide

[0474] MKIILFLALITLATCELYHYQECVRGTTVLLKEPCSSGTYEGNSPFHPLADNKFALTCF

[0475] STQFAFACPDGVKHVYQLRARSVSPKLFIRQEEVQELYSPIFLIVAAIVFITLCFTLKRK TE

[0476] SEQ ID NO:18 amino acid sequence of a SARS-CoV-2 ORF8 polypeptide

[0477] MKFLVFLGIITTVAAFHQECSLQSCTQHQPYVVDDPCPIHFYSKWYIRVGARKSAPLIE

[0478] LCVDEAGSKSPIQYIDIGNYTVSCSPFTINCQEPKLGSLVVRCSFYEDFLEYHDVRVVL DFI

[0479] SEQ ID NO:19 amino acid sequence of a SARS-CoV-2 N polypeptide

[0480] MSDNGPQNQRNAPRITFGGPSDSTGSNQNGERSGARSKQRRPQGLPNNTASWFTALT

[0481] QHGKEDLKFPRGQGVPINTNSSPDDQIGYYRRATRRIRGGDGKMKDLSPRWYFYYL

[0482] GTGPEAGLPYGANKDGIIWVATEGALNTPKDHIGTRNPANNAAIVLQLPQGTTLPKGF YAEGSRGGSQASSRSSSRSRNSSRNSTPGSSRGTSPARMAGNGGDAALALLLLDRLN QLESKMSGKGQQQQGQTVTKKSAAEASKKPRQKRTATKAYNVTQAFGRRGPEQTQ GNFGDQELIRQGTDYKHWPQIAQFAPSASAFFGMSRIGMEVTPSGTWLTYTGAIKLD DKDPNFKDQVILLNKHIDAYKTFPPTEPKKDKKKKADETQALPQRQKKQQTVTLLPA ADLDDFSKQLQQSMSSADSTQA

[0483] SEQ ID NO:20 amino acid sequence of a centralized Hl influenza hemagglutinin polypeptide

[0484] MKAKLLVLLCAFTATDADTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDSHNGK LCKLKGIAPLQLGKCNIAGWILGNPECESLISKRSWSYIVETPNSENGTCYPGDFADY EELREQLSSVSSFERFEIFPKESSWPNHNVTKGVTAACSHAGKSSFYRNLLWLTEKNG SYPKLSKSYVNNKEKEVLVLWGVHHPSNITDQRTLYQNENAYVSVVSSHYNRRFTPE IAKRPKVRGQAGRINYYWTLLEPGDTIIFEANGNLIAPWYAFALSRGFGSGIITSNAPM HECDTKCQTPQGAINSSLPFQNVHPVTIGECPKYVRSTKLRMVTGLRNIPSIQSRGLF GAIAGFIEGGWTGMIDGWYGYHHQNEQGSGYAADQKSTQNAINGITNKVNSVIEKM NTQFTAVGKEFNKLEKRMENLNKKVDDGFLDIWTYNAELLVLLENERTLDFHDSNV KNLYEKVKSQLKNNAKEIGNGCFEFYHKCNNECMESVKNGTYDYPKYSEESKLNRE KIDGVKLESMGVYQILAIYSTVASSLVLLVSLGAISFWMCSNGSLQCRICI

[0485] SEQ ID NO:21 amino acid sequence of a centralized Hl-5 influenza hemagglutinin polypeptide

[0486] MKVKLLILLCTFTATYADTICIGYHANNSTTYVDTITEDNVTVTHATELLESSHNGKL CNLPGVRPLDLGDCSIAGWLLGNPECDLLQNEEWSYIVERSNPANGWCYPGDFPDY EELRSLLASVSSFEKLEIIPEGFSWTNHTQNGGSGACKRGGKSSFFRNLNWLTKKGST YPVLNVSYWNNDNEDKLYIWGVHHPSTDQEQTSLYQNASGYVSVSTSTSQQRIIPNI ASRPTVRGQSGRISFYWTIVAPGDVIVFWSNGNLIAPRYWFKMNAGKSGIMKSDAPI GTCITKCQTPNGAINTSKPFQNVHPITIGECPKYVKSNRLKLATGLRNVPEKQTRGLF GAIAGFIEGGWQGMIDGWYGYHHQNPQGSGYAADLKSTQAAIDGITGKVNIVIEKM NTQFHAVGKEFNELECRIENLNKKVEDGFIDLWTYNAELLVLLENERTLDFHDSNVK NLYEKVRRQLRENAKEIGNGCFEFYHKCDNECMESVRNGTYDYPKYREEAKKNRFQ IKGVKLKSGYKNQILWISFTTVSTLLLVVVLGGIFWCCGNGSLKCRICI

[0487] SEQ ID NO:22 an amino acid sequence of a TcdA polypeptide

[0488] FNLVTGWQTINGKKYYFDINTGAALISYKIINGKHFYFNNDGVMQLGVFKGPDGFEY FAPANTQNNNIEGQ AIVYQ SKFLTLNGKKYYFDQD SKAVTGWRIINNEK YYFNPNNA IAAVGLQVIDNNKYYFNPDTAIISKGWQTVQGSRYYFDTDTAIAFNGYKTIDGKHFYF DSDCVVKIGVFSTSNGFEYFAPANTYNNNIEGQAIVYQSKFLTLNGKKYYFDQNSKA VTGWQTIDSKKYYFNTNTAEAATGWQTIDGKKYYFNTNTAEAATGWQTIDGKKYYF NTNTAIASTGYTIINGKHFYFNTDGIMQIGVFKGPNGFEYFAPANTDANNIEGQAILYQ NEFLTLNGKKYYFGSDSKAVTGWRIINNKKYYFNPNNAIAAIHLCTINNDKYYFSYD GILQNGYITIERNNFYFDANQESKMVTGVFKGPNGFEYFAPANTHNNNIEGQ AIVYQ NKFLTLNGKKYYFDQDSKAVTGWQTIDGKKYYFNLNTAEAATGWQTIDGKKYYFN LNTAEAATGWQTIDGKKYYFNTNTFIASTGYTSINGKHFYFNTDGIMQIGVFKGPNGF EYFAPANTHNNNIEGQAILYQNKFLTLNGKKYYFGSDSKAVTGLRTIDGKKYYFNTN TAVAVTGWQTINGKKYYFNTQTSIASTGYTIISGKHFYFNTDGIMQIGVFKGPDGFEYF APANTDANNIEGQAIRYQNRFLYLHDNIYYFGQNSKAATGWVTIDGNRYYFEPNTAM GANGYKTIDNKNFYFRNGLPQIGVFKGSNGFEYFAPANTDANNIEGQAIRYQNRFLH LLGKIYYFGQNSKAVTGWQTINGKVYYFMPDTAMAAAGGLFEIDGVIYFFGVDGVK APGIYG SEQ ID NO:23 amino acid sequence of a TcdB polypeptide

[0489] NLITGFVTVGDDKYYFNPINGGAASIGETIIDDKNYYFQQSGVLQTGVFSTEDGFKYF APANTLDENLEGEAIDFTGKLIIDENIYYFDDNYRGAVEWKELDGEMHYFSPETGKA FKGLNQIGDYKYYFNSDGVMQKGFVSINDNKHYFDDSGVMKVGYTEIDGKHFYFAE NGEMQIGVFNTEDGFKYFAHHNEDLGNEEGEEISYSGILNFNNKIYYFDDSFTAVVG WKDLEDGSKYYFDEDTAEAYIGLSLINDGQYYFNDDGIMQVGFVTINDKVFYFSDSG IIESGVQNIDDNYFYIDDNGIVQIGVFDTSDGYKYFAPANTVNDNIYGQAVEYSGLVR VGEDVYYFGETYTIETGWIYDMEQESDKYYFNPETKKACKGINLIDDIKYYFDEKGI MRTGLISFENNNYYFNENGEMQFGYINIEDKMFYFGEDGVMQIGVFNTPDGFKYFAH QNTLDENFEGESIQYTGWLDLDEKRYYFTDEYIAATGSVIIDGEEYYFDPDTAQLVISE

[0490] SEQ ID NO:24 nucleic acid sequence that can encode a TcdA polypeptide derived from a C. difficile toxin tttaatctggtgacaggctggcagactatcaatgggaagaaatactatttcgacattaacaccggcgccgctctgatcagctacaagatc attaacgggaaacacttctacttcaacaatgacggagtgatgcagctgggcgtctttaagggccccgatgggttcgagtacttcgcacc tgccaatacacagaacaataacattgaaggacaggccatcgtgtatcagtccaaattcctgactctgaacggcaagaaatactattttga ccaggattctaaggccgtcaccgggtggcgaatcattaataacgagaagtactacttcaaccccaataacgctattgcagccgtgggc ctgcaggtcatcgacaataacaagtactatttcaaccctgatactgccatcatttccaaaggatggcagaccgtgcagggctctcgctac tatttcgacaccgatacagccatcgccttcaacgggtacaagaccatcgacggaaaacatttctattttgactcagattgcgtggtcaaga tcggcgtgttcagcacctccaacggcttcgagtactttgccccagctaacacatacaacaacaacatcgagggccaggccatcgtgta ccagagcaagttcctgaccctgaatggcaagaaatactacttcgaccagaactctaaggcagtcaccgggtggcagacaatcgatagt aagaagtactacttcaacactaacaccgccgaggctgcaactggctggcagaccatcgacgggaagaaatattatttcaatacaaaca ctgccgaagccgctacaggatggcagactattgacggcaagaaatattacttcaacaccaacacagcaatcgcctctacagggtacac tatcattaatggaaagcacttctacttcaacactgatgggatcatgcagattggagtgttcaaaggaccaaatggcttcgagtactttgctc ccgcaaacacagacgccaacaatattgagggccaggctatcctgtatcagaatgaattcctgacactgaacggcaagaaatattatttt gggtctgatagtaaggctgtgactggctggaggatcattaacaataagaagtactatttcaaccccaacaacgcaatcgcagccattca cctgtgcaccattaacaatgacaagtactacttcagctacgacggcatcctgcagaatgggtatatcacaattgagcgcaacaatttcta ctttgacgccaaccaggaatccaagatggtgaccggcgtcttcaaagggcctaatggatttgaatatttcgccccagctaacacacata ataacaatatcgaggggcaggctatcgtgtatcagaataagttcctgaccctgaacggcaagaaatactactttgaccaggatagcaaa gccgtgaccggatggcagacaatcgatggcaagaaatattatttcaatctgaacacagccgaggccgcaactgggtggcagaccatc gatggaaagaaatactacttcaacctgaacactgctgaagccgctaccggatggcagactatcgacgggaagaaatactatttcaatac taacacctttattgcctctaccggatacacaagtatcaatggcaagcacttctacttcaacacggatggaatcatgcagattggcgtgttc aaaggccccaacggatttgaatactttgcacctgccaacactcataataacaatattgaaggccaggctatcctgtaccaaaataagttc ctgaccctgaacgggaagaaatattacttcggatcagacagcaaagccgtgaccggcctgaggacaatcgatgggaagaaatattatt tcaatacgaacactgctgtggcagtcactggatggcagaccattaatggcaagaaatattacttcaacacgcagacaagcatcgcctcc actgggtacaccatcattagcggaaagcacttctacttcaacaccgacggcattatgcagatcggagtgttcaaaggccctgatggattt gagtactttgcccccgctaatacagatgcaaataacattgaaggccaggccatccgataccagaaccggttcctgtatctgcatgacaa tatctactattttggccagaactccaaggcagccacaggctgggtgactatcgatgggaatcggtactatttcgagcctaatacagctat gggggcaaacggatacaagactatcgataacaagaacttctacttccggaatggcctgcctcagatcggggtgtttaagggcagcaa cggattcgagtactttgcaccagccaacaccgacgccaataatattgaaggccaggcaatcagataccagaacaggttcctgcatctg ctgggcaaaatctactacttcggccagaattccaaagcagtgactggctggcagacaatcaacggaaaggtctactacttcatgcctga cacagcaatggctgcagccggcggactgttcgagattgacggcgtgatctacttctttggagtggatggcgtcaaagcacctggaatct acgga

[0491] SEQ ID NO:25 nucleic acid sequence that can encode a TcdB polypeptide derived from a C. difficile toxin aacctgatcactggattcgtgaccgtcggcgacgataagtactacttcaaccctattaacggaggcgctgcatccatcggcgagaccat catcgacgataagaactactacttccagcagagtggggtgctgcagacaggagtcttctcaactgaggacggcttcaagtactttgctc cagcaaataccctggatgaaaacctggagggagaagccattgactttacaggcaagctgatcatcgatgaaaacatctactacttcgac gataactaccgcggagctgtggagtggaaagaactggacggcgagatgcactatttctctccagaaaccggcaaggccttcaaggg gctgaatcagatcggagactacaagtactatttcaacagcgatggcgtgatgcagaaggggtttgtctccatcaatgacaacaaacact acttcgacgatagcggagtgatgaaggtcggctacaccgagattgatggcaaacatttctattttgctgagaatggggaaatgcaaatc ggagtgttcaacacagaagatggcttcaagtactttgcccaccataatgaggacctgggcaacgaggaaggggaggaaatttcctact ctggcatcctgaacttcaacaacaaaatctactatttcgacgatagcttcaccgcagtggtgggatggaaggacctggaggatggaag caaatactattttgacgaggataccgccgaagcttacattggcctgtccctgatcaatgacgggcagtactacttcaacgacgatggcat tatgcaagtggggttcgtcaccatcaacgacaaggtgttctactttagtgattcaggaatcattgagtctggcgtccagaatattgacgat aactacttctatatcgacgataatgggatcgtgcagattggagtcttcgacaccagcgatgggtacaagtattttgcacccgccaacacc gtgaatgacaacatctacggccaggccgtcgagtattcaggcctggtgcgggtcggggaagacgtgtactatttcggcgagacttaca ccattgaaacagggtggatctatgacatggagcaagaaagtgataagtactatttcaatcctgagactaagaaagcctgcaaaggcatc aacctgattgacgatatcaagtactacttcgatgagaagggaatcatgagaaccggcctgatcagcttcgaaaacaataactactacttc aacgagaacggggaaatgcagttcggatacatcaacatcgaggacaagatgttctacttcggggaagatggagtgatgcagatcgga gtctttaacacacccgacggcttcaaatactttgcccaccagaatactctggatgagaacttcgagggggaatctatccagtacaccgg atggctggacctggatgagaagaggtactatttcaccgacgagtacatcgccgctacaggcagtgtgattatcgacggcgaggagtat tacttcgatcccgacaccgctcagctggtcatctcagag

[0492] SEQ ID NO:26 nucleic acid sequence that can encode a SC-Ad-Spike virus cgccatcatcaataatataccttattttggattgaagccaatatgataatgagggggtggagtttgtgacgtggcgcggggcgtgggaac ggggcgggtgacgtagtagtgtggcggaagtgtgatgttgtaagtgtggcggaacacatgtaagcgccggatgtggtaaaagtgacg tttttggtgtgcgccggtgtacacgggaagtgacaattttcgcgcggttttaggcggatgttgtagtaaatttgggcgtaaccaagtaatat ttggccattttcgcgggaaaactgaataagaggaagtgaaatctgaataattctgtgttactcatagcgcgtaatatttgtctagggccgc ggggactttgaccgtttacgtggagactcgcccaggtgtttttctcaggtgttttccgcgttccgggtcaaagttggcgttttattattatagt cagctgacgcgcagtgtatttatacccggtgagttcctcaagaggccactcttgagtgccagcgagtagagttttctcctccgagccgct ccgacaccgggactgaaaatgagacatattatctgccacggaggtgttattaccgaagaaatggccgccagtcttttggaccagctgat cgaagaggtactggctgataatcttccacctcctagccattttgaaccacctacccttcacgaactgtatgatttagacgtgacggccccc gaagatcccaacgaggaggcggtttcgcagatttttcccgagtctgtaatgttggcggtgcaggaagggattgacttattcacttttccgc cggcgcccggttctccggagccgcctcacctttcccggcagcccgagcagccggagcagagagccttgggtccggtttctatgcca aaccttgtgccggaggtgatcgatcttacctgccacgaggctggctttccacccagtgacgacgaggatgaagagggtgaggagtttg tgttagattatgtggagcaccccgggcacggttgcaggtcttgtcattatcaccggaggaatacgggggacccagatattatgtgttcgc tttgctatatgaggacctgtggcatgtttgtctacagtaagtgaaaaattatgggcagtgggtgatagagtggtgggtttggtgtggtaatt tttttttaatttttacagttttgtggtttaaagaattttgtattgtgattttttaaaaggtcctgtgtctgaacctgagcctgagcccgagccagaa ccggagcctgcaagacctacccggcgtcctaaattggtgcctgctatcctgagacgcccgacatcacctgtgtctagagaatgcaata gtagtacggatagctgtgactccggtccttctaacacacctcctgagatacacccggtggtcccgctgtgccccattaaaccagttgcc gtgagagttggtgggcgtcgccaggctgtggaatgtatcgaggacttgcttaacgagtctgggcaacctttggacttgagctgtaaacg ccccaggccataaggtgtaaacctgtgattgcgtgtgtggttaacgcctttgtttgctgaatgagttgatgtaagtttaataaagggtgaga taatgtttaacttgcatggcgtgttaaatggggcggggcttaaagggtatataatgcgccgtgggctaatcttggttacatctgacctcatg gaggcttgggagtgtttggaagatttttctgctgtgcgtaacttgctggaacagagctctaacagtacctcttggttttggaggtttctgtgg ggctcctcccaggcaaagttagtctgcagaattaaggaggattacaagtgggaatttgaagagcttttgaaatcctgtggtgagctgtttg attctttgaatctgggtcaccaggcgcttttccaagagaaggtcatcaagactttggatttttccacaccggggcgcgctgcggctgctgt tgcttttttgagttttataaaggataaatggagcgaagaaacccatctgagcggggggtacctgctggattttctggccatgcatctgtgg agagcggtggtgagacacaagaatcgcctgctactgttgtcttccgtccgcccggcaataataccgacggaggagcaacagcagga ggaagccaggcggcggcggcggcaggagcagagcccatggaacccgagagccggcctggaccctcgggaatgaatgttgtaca ggtggctgaactgtttccagaactgagacgcattttaaccattaacgaggatgggcaggggctaaagggggtaaagagggagcggg gggcttctgaggctacagaggaggctaggaatctaacttttagcttaatgaccagacaccgtcctgagtgtgttacttttcagcagattaa ggataattgcgctaatgagcttgatctgctggcgcagaagtattccataaagcagctgaccacttactggctgcagccaggggatgattt tgaggaggctattagggtatatgcaaaggtggcacttaggccagattgcaagtacaagattagcaaacttgtaaatatcaggaattgttg ctacatttctgggaacggggccgaggtggagatagatacggaggatagggtggcctttagatgtagcatgataaatatgtggccggg ggtgcttggcatggacggggtggttattatgaatgtgaggtttactggtcccaattttagcggtacggttttcctggccaataccaatcttat cctacacggtgtaagcttctatgggtttaacaatacctgtgtggaagcctggaccgatgtaagggttcggggctgtgccttttactgctgc tggaagggggtggtgtgtcgccccaaaagcagggcttcaattaagaaatgcctgtttgaaaggtgtaccttgggtatcctgtctgaggg taactccagggtgcgccacaatgtggcctccgactgtggttgctttatgctagtgaaaagcgtggctgtgattaagcataacatggtgtgt ggcaactgcgaggacagggcctctcagatgctgacctgctcggacggcaactgtcacttgctgaagaccattcacgtagccagccac tctcgcaaggcctggccagtgtttgagcacaacatactgacccgctgttccttgcatttgggtaacaggaggggggtgttcctaccttac caatgcaatttgagtcacactaagatattgcttgagcccgagagcatgtccaaggtgaacctgaacggggtgtttgacatgaccatgaa gatctggaaggtgctgaggtacgatgagacccgcaccaggtgcagaccctgcgagtgtggcggtaaacatattaggaaccagcctgt gatgctggatgtgaccgaggagctgaggcccgatcacttggtgctggcctgcacccgcgctgagtttggctctagcgatgaagataca gattgaggtactgaaatgtgtgggcgtggcttaagggtgggaaagaatatataaggtgggggtctcatgtagttttgtatctgttttgcag cagccgccgccatgagcgccaactcgtttgatggaagcattgtgagctcatatttgacaacgcgcatgcccccatgggccggggtgc gtcagaatgtgatgggctccagcattgatggtcgccccgtcctgcccgcaaactctactaccttgacctacgagaccgtgtctggaacg ccgttggagactgcagcctccgccgccgcttcagccgctgcagccaccgcccgcgggattgtgactgactttgctttcctgagcccgc ttgcaagcagtgcagcttcccgttcatccgcccgcgatgacaagttgacggctcttttggcacaattggattctttgacccgggaacttaa tgtcgtttctcagcagctgttggatctgcgccagcaggtttctgccctgaaggcttcctcccctcccaatgcggtttaaaacataaataaaa accagactctgtttggatttggatcaagcaagtgtcttgctgtctttatttaggggttttgcgcgcgcggtaggcccgggaccagcggtct cggtcgttgagggtcctgtgtattttttccaggacgtggtaaaggtgactctggatgttcagatacatgggcataagcccgtctctggggt ggaggtagcaccactgcagagcttcatgctgcggggtggtgttgtagatgatccagtcgtagcaggagcgctgggcgtggtgcctaa aaatgtctttcagtagcaagctgattgccaggggcaggcccttggtgtaagtgtttacaaagcggttaagctgggatgggtgcatacgt ggggatatgagatgcatcttggactgtatttttaggttggctatgttcccagccatatccctccggggattcatgttgtgcagaaccaccag cacagtgtatccggtgcacttgggaaatttgtcatgtagcttagaaggaaatgcgtggaagaacttggagacgcccttgtgacctccaa gattttccatgcattcgtccataatgatggcaatgggcccacgggcggcggcctgggcgaagatatttctgggatcactaacgtcatagt tgtgttccaggatgagatcgtcataggccatttttacaaagcgcgggcggagggtgccagactgcggtataatggttccatccggccca ggggcgtagttaccctcacagatttgcatttcccacgctttgagttcagatggggggatcatgtctacctgcggggcgatgaagaaaac cgtttccggggtaggggagatcagctgggaagaaagcaggttcctaagcagctgcgacttaccgcagccggtgggcccgtaaatca cacctattaccggctgcaactggtagttaagagagctgcagctgccgtcatccctgagcaggggggccacttcgttaagcatgtccctg acttgcatgttttccctgaccaaatccgccagaaggcgctcgccgcccagcgatagcagttcttgcaaggaagcaaagtttttcaacgg tttgaggccgtccgccgtaggcatgcttttgagcgtttgaccaagcagttccaggcggtcccacagctcggtcacgtgctctacggcat ctcgatccagcatatctcctcgtttcgcgggttggggcggctttcgctgtacggcagtagtcggtgctcgtccagacgggccagggtca tgtctttccacgggcgcagggtcctcgtcagcgtagtctgggtcacggtgaaggggtgcgctccgggttgcgcgctggccagggtgc gcttgaggctggtcctgctggtgctgaagcgctgccggtcttcgccctgcgcgtcggccaggtagcatttgaccatggtgtcatagtcc agcccctccgcggcgtggcccttggcgcgcagcttgcccttggaggaggcgccgcacgaggggcagtgcagacttttaagggcgt agagcttgggcgcgagaaataccgattccggggagtaggcatccgcgccgcaggccccgcagacggtctcgcattccacgagcca ggtgagctctggccgttcggggtcaaaaaccaggtttcccccatgctttttgatgcgtttcttacctctggtttccatgagccggtgtccac gctcggtgacgaaaaggctgtccgtgtccccgtatacagacttgagaggcctgtcctcgagcggtgttccgcggtcctcctcgtataga aactcggaccactctgagacgaaggctcgcgtccaggccagcacgaaggaggctaagtgggaggggtagcggtcgttgtccacta gggggtccactcgctccagggtgtgaagacacatgtcgccctcttcggcatcaaggaaggtgattggtttataggtgtaggccacgtg accgggtgttcctgaaggggggctataaaagggggtgggggcgcgttcgtcctcactctcttccgcatcgctgtctgcgagggccag ctgttggggtgagtactccctctcaaaagcgggcatgacttctgcgctaagattgtcagtttccaaaaacgaggaggatttgatattcacc tggcccgcggtgatgcctttgagggtggccgcgtccatctggtcagaaaagacaatctttttgttgtcaagcttggtggcaaacgaccc gtagagggcgttggacagcaacttggcgatggagcgcagggtttggtttttgtcgcgatcggcgcgctccttggccgcgatgtttagct gcacgtattcgcgcgcaacgcaccgccattcgggaaagacggtggtgcgctcgtcgggcactaggtgcacgcgccaaccgcggtt gtgcagggtgacaaggtcaacgctggtggctacctctccgcgtaggcgctcgttggtccagcagaggcggccgcccttgcgcgagc agaatggcggtagtgggtctagctgcgtctcgtccggggggtctgcgtccacggtaaagaccccgggcagcaggcgcgcgtcgaa gtagtctatctgcatcctgcaagtctagcgcctgctgccatgcgcgggcggcaagcgcgcgctcgtatgggttgagtgggggaccc catggcatggggtgggtgagcgcggaggcgtacatgccgcaaatgtcgtaaacgtagaggggctctctgagtattccaagatatgta gggtagcatcttccaccgcggatgctggcgcgcacgtaatcgtatagttcgtgcgagggagcgaggaggtcgggaccgaggttgct acgggcgggctgctctgctcggaagactatctgcctgaagatggcatgtgagttggatgatatggttggacgctggaagacgttgaag ctggcgtctgtgagacctaccgcgtcacgcacgaaggaggcgtaggagtcgcgcagcttgttgaccagctcggcggtgacctgcac gtctagggcgcagtagtccagggtttccttgatgatgtcatacttatcctgtcccttttttttccacagctcgcggttgaggacaaactcttc gcggtctttccagtactcttggatcggaaacccgtcggcctccgaacggtaagagcctancatgtagaactggttgacggcctggtag gcgcagcatcccttttctacgggtagcgcgtatgcctgcgcggccttccggagcgaggtgtgggtgagcgcaaaggtgtccctaacc atgactttgaggtactggtatttgaagtcagtgtcgtcgcatccgccctgctcccagagcaaaaagtccgtgcgctttttggaacgcggg tttggcagggcgaaggtgacatcgttgaagagtatctttcccgcgcgaggcataaagttgcgtgtgatgcggaagggtcccggcacct cggaacggttgttaattacctgggcggcgagcacgatctcgtcaaagccgttgatgttgtggcccacaatgtaaagttccaagaagcg cgggatgcccttgatggaaggcaattttttaagttcctcgtaggtgagctcttcaggggagctgagcccgtgctctgaaagggcccagt ctgcaagatgagggttggaagcgacgaatgagctccacaggtcacgggccattagcatttgcaggtggtcgcgaaaggtcctaaact ggcgacctatggccattttttctggggtgatgcagtagaaggtaagcgggtcttgttcccagcggtcccatccaaggtccgcggctagg tctcgcgcggcggtcactagaggctcatctccgccgaacttcatgaccagcatgaagggcacgagctgcttcccaaaggcccccatc caagtataggtctctacatcgtaggtgacaaagagacgctcggtgcgaggatgcgagccgatcgggaagaactggatctcccgcca ccagttggaggagtggctgttgatgtggtgaaagtagaagtccctgcgacgggccgaacactcgtgctggcttttgtaaaaacgtgcg cagtactggcagcggtgcacgggctgtacatcctgcacgaggttgacctgacgaccgcgcacaaggaagcagagtgggaatttgag cccctcgcctggcgggtttggctggtggtcttctacttcggctgcttgtccttgaccgtctggctgctcgaggggagttacggtggatcg gaccaccacgccgcgcgagcccaaagtccagatgtccgcgcgcggcggtcggagcttgatgacaacatcgcgcagatgggagct gtccatggtctggagctcccgcggcgtcaggtcaggcgggagctcctgcaggtttacctcgcatagccgggtcagggcgcgggcta ggtccaggtgatacctgatttccaggggctggttggtggcggcgtcgatggcttgcaagaggccgcatccccgcggcgcgactacg gtaccgcgcggcgggcggtgggccgcgggggtgtccttggatgatgcatctaaaagcggtgacgcgggcgggcccccggaggta gggggggctcgggacccgccgggagagggggcaggggcacgtcggcgccgcgcgcgggcaggagctggtgctgcgcgcgg aggttgctggcgaacgcgacgacgcggcggttgatctcctgaatctggcgcctctgcgtgaagacgacgggcccggtgagcttgaa cctgaaagagagttcgacagaatcaatttcggtgtcgttgacggcggcctggcgcaaaatctcctgcacgtctcctgagttgtcttgata ggcgatctcggccatgaactgctcgatctcttcctcctggagatctccgcgtccggctcgctccacggtggcggcgaggtcgttggag atgcgggccatgagctgcgagaaggcgttgaggcctccctcgttccagacgcggctgtagaccacgcccccttcggcatcgcgggc gcgcatgaccacctgcgcgagattgagctccacgtgccgggcgaagacggcgtagtttcgcaggcgctgaaagaggtagttgagg gtggtggcggtgtgttctgccacgaagaagtacataacccagcgccgcaacgtggattcgttgatatcccccaaggcctcaaggcgct ccatggcctcgtagaagtccacggcgaagttgaaaaactgggagttgcgcgccgacacggttaactcctcctccagaagacggatga gctcggcgacagtgtcgcgcacctcgcgctcaaaggctacaggggcctcttcttcttcttcaatctcctcttccataagggcctccccttc ttcttcttctggcggcggtgggggaggggggacacggcggcgacgacggcgcaccgggaggcggtcgacaaagcgctcgatcat ctccccgcggcgacggcgcatggtctcggtgacggcgcggccgttctcgcgggggcgcagttggaagacgccgcccgtcatgtcc cggttatgggttggcggggggctgccgtgcggcagggatacggcgctaacgatgcatctcaacaattgttgtgtaggtactccgccac cgagggacctgagcgagtccgcatcgaccggatcggaaaacctctcgagaaaggcgtctaaccagtcacagtcgcaaggtaggct gagcaccgtggcgggcggcagcgggcggcggtcggggttgtttctggcggaggtgctgctgatgatgtaattaaagtaggcggtctt gagacggcggatggtcgacagaagcaccatgtccttgggtccggcctgctgaatgcgcaggcggtcggccatgccccaggcttcgt tttgacatcggcgcaggtctttgtagtagtcttgcatgagcctttctaccggcacttcttcttctccttcctcttgtcctgcatctcttgcatctat cgctgcggcggcggcggagtttggccgtaggtggcgccctcttcctcccatgcgtgtgaccccgaagcccctcatcggctgaagca gggccaggtcggcgacaacgcgctcggctaatatggcctgctgcacctgcgtgagggtagactggaagtcgtccatgtccacaaag cggtggtatgcgcccgtgttgatggtgtaagtgcagttggccataacggaccagttaacggtctggtgacccggctgcgagagctcgg tgtacctgagacgcgagtaagcccttgagtcaaagacgtagtcgttgcaagtccgcaccaggtactggtatcccaccaaaaagtgcgg cggcggctggcggtagaggggccagcgtagggtggccggggctccgggggcgaggtcttccaacataaggcgatgatatccgta gatgtacctggacatccaggtgatgccggcggcggtggtggaggcgcgcggaaagtcacggacgcggttccagatgttgcgcagc ggcaaaaagtgctccatggtcgggacgctctggccggtcaggcgcgcgcagtcgttgacgctctagaccgtgcaaaaggagagcct gtaagcgggcactcttccgtggtctggtggataaattcgcaagggtatcatggcggacgaccggggttcgaaccccggatccggccg tccgccgtgatccatgcggttaccgcccgcgtgtcgaacccaggtgtgcgacgtcagacaacgggggagcgctccttttggcttcctt ccaggcgcggcggatgctgcgctagcttttttggccactggccgcgcgcggcgtaagcggttaggctggaaagcgaaagcattaagt ggctcgctccctgtagccggagggttattttccaagggttgagtcgcgggacccccggttcgagtctcgggccggccggactgcggc gaacgggggtttgcctccccgtcatgcaagaccccgcttgcaaattcctccggaaacagggacgagccccttttttgcttttcccagatg catccggtgctgcggcagatgcgcccccctcctcagcagcggcaagagcaagagcagcggcagacatgcagggcaccctcccct cctcctaccgcgtcaggaggggcgacatccgcggttgacgcggcagcagatggtgattacgaacccccgcggcgccgggcccgg cactacctggactggaggagggcgagggcctggcgcggctaggagcgccctctcctgagcggtacccaagggtgcagctgaagc gtgatacgcgtgaggcgtacgtgccgcggcagaacctgtttcgcgaccgcgagggagaggagcccgaggagatgcgggatcgaa agttccacgcagggcgcgagctgcggcatggcctgaatcgcgagcggttgctgcgcgaggaggactttgagcccgacgcgcgaac cgggattagtcccgcgcgcgcacacgtggcggccgccgacctggtaaccgcatacgagcagacggtgaaccaggagattaactttc aaaaaagctttaacaaccacgtgcgtacgcttgtggcgcgcgaggaggtggctataggactgatgcatctgtgggactttgtaagcgc gctggagcaaaacccaaatagcaagccgctcatggcgcagctgttccttatagtgcagcacagcagggacaacgaggcattcaggg atgcgctgctaaacatagtagagcccgagggccgctggctgctcgatttgataaacatcctgcagagcatagtggtgcaggagcgca gcttgagcctggctgacaaggtggccgccatcaactattccatgcttagcctgggcaagttttacgcccgcaagatataccatacccctt acgttcccatagacaaggaggtaaagatcgaggggttctacatgcgcatggcgctgaaggtgcttaccttgagcgacgacctgggcg tttatcgcaacgagcgcatccacaaggccgtgagcgtgagccggcggcgcgagctcagcgaccgcgagctgatgcacagcctgca aagggccctggctggcacgggcagcggcgatagagaggccgagtcctactttgacgcgggcgctgacctgcgctgggccccaag ccgacgcgccctggaggcagctggggccggacctgggctggcggtggcacccgcgcgcgctggcaacgtcggcggcgtggag gaatatgacgaggacgatgagtacgagccagaggacggcgagtactaagcggtgatgtttctgatcagtcgcggccgcgatatcgct agcgaagttcctattctctagaaagtataggaacttcggatcctctagagtcgaaaaaaaaaaagcatgatgcaaaataaaaaactcac caaggccatggcaccgagcgttggttttcttgtattccccttagtatgcggcgcgcggcgatgtatgaggaaggtcctcctccctcctac gagagtgtggtgagcgcggcgccagtggcggcggcgctgggttctcccttcgatgctcccctggacccgccgtttgtgcctccgcgg tacctgcggcctaccggggggagaaacagcatccgttactctgagttggcacccctattcgacaccacccgtgtgtacctggtggaca acaagtcaacggatgtggcatccctgaactaccagaacgaccacagcaactttctgaccacggtcattcaaaacaatgactacagccc gggggaggcaagcacacagaccatcaatcttgacgaccggtcgcactggggcggcgacctgaaaaccatcctgcataccaacatg ccaaatgtgaacgagttcatgtttaccaataagtttaaggcgcgggtgatggtgtcgcgcttgcctactaaggacaatcaggtggagct gaaatacgagtgggtggagttcacgctgcccgagggcaactactccgagaccatgaccatagaccttatgaacaacgcgatcgtgga gcactacttgaaagtgggcagacagaacggggttctggaaagcgacatcggggtaaagtttgacacccgcaacttcagactggggtt tgaccccgtcactggtcttgtcatgcctggggtatatacaaacgaagccttccatccagacatcattttgctgccaggatgcggggtgga cttcacccacagccgcctgagcaacttgttgggcatccgcaagcggcaacccttccaggagggctttaggatcacctacgatgatctg gagggtggtaacattcccgcactgttggatgtggacgcctaccaggcgagcttgaaagatgacaccgaacagggcgggggtggcg caggcggcagcaacagcagtggcagcggcgcggaagagaactccaacgcggcagccgcggcaatgcagccggtggaggacat gaacgatcatgccattcgcggcgacacctttgccacacgggctgaggagaagcgcgctgaggccgaagcagcggccgaagctgc cgcccccgctgcgcaacccgaggtcgagaagcctcagaagaaaccggtgatcaaacccctgacagaggacagcaagaaacgcag ttacaacctaataagcaatgacagcaccttcacccagtaccgcagctggtaccttgcatacaactacggcgaccctcagaccggaatc cgctcatggaccctgctttgcactcctgacgtaacctgcggctcggagcaggtctactggtcgttgccagacatgatgcaagaccccgt gaccttccgctccacgcgccagatcagcaactttccggtggtgggcgccgagctgttgcccgtgcactccaagagcttctacaacgac caggccgtctactcccaactcatccgccagtttacctctctgacccacgtgttcaatcgctttcccgagaaccagattttggcgcgcccg ccagcccccaccatcaccaccgtcagtgaaaacgttcctgctctcacagatcacgggacgctaccgctgcgcaacagcatcggagg agtccagcgagtgaccattactgacgccagacgccgcacctgcccctacgtttacaaggccctgggcatagtctcgccgcgcgtcct atcgagccgcactttttgagcaagcatgtccatccttatatcgcccagcaataacacaggctggggcctgcgcttcccaagcaagatgtt tggcggggccaagaagcgctccgaccaacacccagtgcgcgtgcgcgggcactaccgcgcgccctggggcgcgcacaaacgcg gccgcactgggcgcaccaccgtcgatgacgccatcgacgcggtggtggaggaggcgcgcaactacacgcccacgccgccgcca gtgtccaccgtggacgcggccattcagaccgtggtgcgcggagcccggcgctacgctaaaatgaagagacggcggaggcgcgta gcacgtcgccaccgccgccgacccggcactgccgcccaacgcgcggcggcggccctgcttaaccgcgcacgtcgcaccggccg acgggcggccatgcgagccgctcgaaggctggccgcgggtattgtcactgtgccccccaggtccaggcgacgagcggccgccgc agcagccgcggccattagtgttatgactcagggtcgcaggggcaacgtgtactgggtgcgcgactcggttagcggcctgcgcgtgc ccgtgcgcacccgccccccgcgcaactagattgcaataaaaaactacttagactcgtactgttgtatgtatccagcggcggcggcgcg catcgaagctatgtccaagcgcaaaatcaaagaagagatgctccaggtcatcgcgccggagatctatggccccccgaagaaggaag agcaggattacaagccccgaaagctaaagcgggtcaaaaagaaaaagaaagatgatgatgatgatgaacttgacgacgaggtggaa ctgttgcacgcgaccgcgcccaggcgacgggtacagtggaaaggtcgacgcgtaagacgtgttttgcgacccgg caeca ccgtagt ctttacgcccggtgagcgctccacccgcacctacaagcgcgtgtatgatgaggtgtacggcgacgaggacctgcttgagcaggcca acgagcgcctcggggagtttgcctacggaaagcggcataaggacatgctggcgttgccgctggacgagggcaacccaacacctag cctaaagcccgtgacactgcagcaggtgctgcccgcgcttgcaccgtccgaagaaaagcgcggcctaaagcgcgagtctggtgact tggcacccaccgtgcagctgatggtacccaagcgtcagcgactggaagatgtcttggaaaaaatgaccgtggagcctgggctggag cccgaggtccgcgtgcggccaatcaagcaggtggcaccgggactgggcgtgcagaccgtggacgttcagatacccaccaccagta gcactagtattgccactgccacagagggcatggagacacaaacgtccccggttgcctcggcggtggcagatgccgcggtgcaggc ggccgctgcggccgcgtccaagacctctacggaggtgcaaacggacccgtggatgtttcgtgtttcagccccccggcgtccgcgcc gttcaaggaagtacggcgccgccagcgcgctactgcccgaatatgccctacatccttccatcgcgcctacccccggctatcgtggcta cacctaccgccccagaagacgagcaactacccgacgccgaaccaccactggaacccgccgccgccgtcgccgtcgccagcccgt gctggccccgatttccgtgcgcagggtggctcgcgaaggaggcaggaccctggtgctgccaacagcgcgctaccaccccagcatc gtttaaaagccggtctttgtggttcttgcagatatggccctcacctgccgcctccgtttcccggtgccgggattccgaggaagaatgcac cgtaggaggggcatggccggccacggcctgacgggcggcatgcgtcgtgcgcaccaccggcggcggcgcgcgtcgcaccgtcg catgcgcgccggtatcctgcccctccttattccactgatcgccgcggcgattggcgccgtgcccggaattgcatccgtggccttgcag gcgcagagacactgattaaaaacaagttacatgtggaaaaatcaaaataaaagtctggactctcacgctcgcttggtcctgtaactatttt gtagaatggaagacatcaactttgcgtcactggccccgcgacacggctcgcgcccgttcatgggaaactggcaagatatcgg caeca gcaatatgagcggtggcgccttcagctggggctcgctgtggagcggcattaaaaatttcggttccgccgttaagaactatggcagcaa agcctggaacagcagcacaggccagatgctgagggacaagttgaaagagcaaaatttccaacaaaaggtggtagatggcctggcct ctggcattagcggggtggtggacctggccaaccaggcagtgcaaaataagattaacagtaagcttgatccccgccctcccgtagagg agcctccaccggccgtggagacagtgtctccagaggggcgtggcgaaaagcgtccgcgacccgacagggaagaaactctggtga cgcaaatagacgagcctccctcgtacgaggaggcactaaagcaaggcctgcccaccacccgtcccatcgcgcccatggctaccgg agtgctgggccagcacacacccgtaacgctggacctgcctccccccgccgacacccagcagaaacctgtgctgccaggcccgtcc gccgttgttgtaacccgtcctagccgcgggtccctgcgccgcgccgccagcggtccgcgatcgttgcggcccgtagccagtggcaa ctggcaaagcacactgaacagcatcgtgggtttgggggtgcaatccctgaagcgccgacgatgcttctgatagctaacgtgtcgtatgt gtgtcatgtatgcgtccatgtcgccgccagaggagctgctgagccgccgcgcgcccgctttccaagatggctaccccttcgatgatgc cgcagtggtcttacatgcacatctcgggccaggacgcctcggagtacctgagccccgggctggtgcagttcgcccgcgccaccgag acgtacttcagcctgaataacaagtttagaaaccccacggtggcgcctacgcacgacgtgaccacagaccggtctcagcgtttgacgc tgcggttcatccccgtggaccgcgaggatactgcgtactcgtacaaggcgcggttcaccctagctgtgggtgataaccgtgtgctaga catggcttccacgtactttgacatccgcggcgtgctggacaggggccctacttttaagccctactctggcactgcctacaacgcactgg cccccaagggtgcccccaactcgtgcgagtgggaacaaaatgaaactgcacaagtggatgctcaagaacttgacgaagaggagaat gaagccaatgaagctcaggcgcgagaacaggaacaagctaagaaaacccatgtatatgcccaggctccactgtccggaataaaaat aactaaagaaggtctacaaataggaactgccgacgccacagtagcaggtgccggcaaagaaattttcgcagacaaaacttttcaacct gaaccacaagtaggagaatctcaatggaacgaagcggatgccacagcagctggtggaagggttcttaaaaagacaactcccatgaa accctgctatggctcatacgctagacccaccaattccaacggcggacagggcgttatggttgaacaaaatggtaaattggaaagtcaa gtcgaaatgcaatttttttccacatccacaaatgccacaaatgaagttaacaatatacaaccaacagttgtattgtacagcgaagatgtaaa catggaaactccagatactcatctttcttataaacctaaaatgggggataaaaatgccaaagtcatgcttggacaacaagcaatgccaaa cagaccaaattacattgcttttagagacaattttattggtctcatgtattacaacagcacaggtaacatgggtgtccttgctggtcaggcatc gcagttgaacgctgttgtagatttgcaagacagaaacacagagctgtcctaccagcttttgcttgattcaattggcgacagaacaagata cttttcaatgtggaatcaagctgttgacagctatgatccagatgtcagaattattgagaaccatggaactgaggatgagttgccaaattatt gctttcctcttggtggaattgggattactgacacttttcaagctgttaaaacaactgctgctaacggggaccaaggcaatactacctggca aaaagattcaacatttgcagaacgcaatgaaataggggtgggaaataactttgccatggaaattaacctgaatgccaacctatggagaa atttcctttactccaatattgcgctgtacctgccagacaagctaaaatacaaccccaccaatgtggaaatatctgacaaccccaacaccta cgactacatgaacaagcgagtggtggctcctgggcttgtagactgctacattaaccttggggcgcgctggtctctggactacatggaca acgttaatccctttaaccacccccgccatgcgggcctgcgttaccgctccatgttgttgggaaacggccgctacgtgccctttcacattc aggtgccccaaaagttttttgccattaaaaacctcctcctcctgccaggctcatacacatatgaatggaacttcaggaaggatgttaacat ggttctgcagagctctctgggaaacgaccttagagttgacggggctagcattaagtttgacagcatttgtctttacgccaccttcttcccca tggcccacaacacggcctccacgctggaagccatgctcagaaatgacaccaacgaccagtcctttaatgactacctttccgccgccaa catgctatatcccatacccgccaacgccaccaacgtgcccatctccatcccatcgcgcaactgggcagcatttcgcggttgggccttca cacgcttgaagacaaaggaaaccccttccctgggatcaggctacgacccttactacacctactctggctccataccataccttgacgga accttctatcttaatcacacctttaagaaggtggccattacttttgactcttctgttagctggccgggcaacgaccgcctgcttactcccaat gagtttgagattaagcgctcagttgacggggagggctataacgtagctcagtgcaacatgacaaaggactggttcctagtgcagatgtt ggccaactacaatattggctaccagggcttctacattccagaaagctacaaagaccgcatgtactcgttcttcagaaacttccagcccat gagccggcaagtggtggacgatactaaatacaaagattatcagcaggttggaattatccaccagcataacaactcaggcttcgtaggct acctcgctcccaccatgcgcgagggacaagcttaccccgctaatgttccctacccactaataggcaaaaccgcggttgatagtattacc cagaaaaagtttctttgcgaccgcaccctgtggcgcatccccttctccagtaactttatgtccatgggtgcgctcacagacctgggccaa aaccttctctacgcaaactccgcccacgcgctagacatgacctttgaggtggatcccatggacgagcccacccttctttatgttttgtttga agtctttgacgtggtccgtgtgcaccagccgcaccgcggcgtcatcgagaccgtgtacctgcgcacgcccttctcggccggcaacgc cacaacataaagaagcaagcaacatcaacaacagctgccgccatgggctccagtgagcaggaactgaaagccattgtcaaagatctt ggttgtgggccatattttttgggcacctatgacaagcgcttcccaggctttgtttccccacacaagctcgcctgcgccatagttaacacgg ccggtcgcgagactgggggcgtacactggatggcctttgcctggaacccgcgctcaaaaacatgctacctctttgagccctttggctttt ctgaccaacgtctcaagcaggtttaccagtttgagtacgagtcactcctgcgccgtagcgccattgcctcttcccccgaccgctgtataa cgctggaaaagtccacccaaagcgtgcaggggcccaactcggccgcctgtggcctattctgctgcatgtttctccacgcctttgccaac tggccccaaactcccatggatcacaaccccaccatgaaccttattaccggggtacccaactccatgcttaacagtccccaggtacagc ccaccctgcgccgcaaccaggaacagctctacagcttcctggagcgccactcgccctacttccgcagccacagtgcgcaaattagga gcgccacttctttttgtcacttgaaaaacatgtaaaaataatgtactaggagacactttcaataaaggcaaatgtttttatttgtacactctcg ggtgattatttacccccacccttgccgtctgcgccgtttaaaaatcaaaggggttctgccgcgcatcgctatgcgccactggcagggac acgttgcgatactggtgtttagtgctccacttaaactcaggcacaaccatccgcggcagctcggtgaagttttcactccacaggctgcgc accatcaccaacgcgtttagcaggtcgggcgccgatatcttgaagtcgcagttggggcctccgccctgcgcgcgcgagttgcgatac acagggttacagcactggaacactatcagcgccgggtggtgcacgctggccagcacgctcttgtcggagatcanatccgcgtccag gtcctccgcgttgctcagggcgaacggagtcaactttggtagctgccttcccaaaaagggtgcatgcccaggctttgagttgcactcgc accgtagtggcatcagaaggtgaccgtgcccagtctgggcgttaggatacagcgcctgcatgaaagccttgatctgcttaaaagccac ctgagcctttgcgccttcagagaagaacatgccgcaagacttgccggaaaactgattggccggacaggccgcgtcatgcacgcagc accttgcgtcggtgttggagatctgcaccacatttcggccccaccggttcttcacgatcttggccttgctagactgctccttcagcgcgcg ctgcccgttttcgctcgtcacatccatttcaatcacgtgctccttatttatcataatgctcccgtgtagacacttaagctcgccttcgatctca gcgcagcggtgcagccacaacgcgcagcccgtgggctcgtggtgcttgtaggttacctctgcaaacgactgcaggtacgcctgcag gaatcgccccatcatcgtcacaaaggtcttgttgctggtgaaggtcagctgcaacccgcggtgctcctcgtttagccaggtcttgcatac ggccgccagagcttccacttggtcaggcagtagcttgaagtttgcctttagatcgttatccacgtggtacttgtccatcaacgcgcgcgc agcctccatgcccttctcccacgcagacacgatcggcaggctcagcgggtttatcaccgtgctttcactttccgcttcactggactcttcc ttttcctcttgcatccgcataccccgcgccactgggtcgtcttcattcagccgccgcaccgtgcgcttacctcccttgccgtgcttgattag caccggtgggttgctgaaacccaccatttgtagcgccacatcttctctttcttcctcgctgtccacgatcacctctggggatggcgggcg ctcgggcttgggagaggggcgcttctttttctttttggacgcaatggccaaatccgccgtcgaggtcgatggccgcgggctgggtgtgc gcggcaccagcgcatcttgtgacgagtcttcttcgtcctcggactcgagacgccgcctcagccgcttttttgggggcgcgcggggag gcggcggcgacggcgacggggacgagacgtcctccatggttggtggacgtcgcgccgcaccgcgtccgcgctcgggggtggttt cgcgctgctcctcttcccgactggccatttccttctcctataggcagaaaaagatcatggagtcagtcgagaaggaggacagcctaacc gccccctttgagttcgccaccaccgcctccaccgatgccgccaacgcgcctaccaccttccccgtcgaggcacccccgcttgaggag gaggaagtgattatcgagcaggacccaggttttgtaagcgaagacgacgaagatcgctcagtaccaacagaggataaaaagcaaga ccaggacgacgcagaggcaaacgaggaacaagtcgggcggggggaccaaaggcatggcgactacctagatgtgggagacgac gtgctgttgaagcatctgcagcgccagtgcgccattatctgcgacgcgttgcaagagcgcagcgatgtgcccctcgccatagcggat gtcagccttgcctacgaacgccacctgttctcaccgcgcgtaccccccaaacgccaagaaaacggcacatgcgagcccaacccgcg cctcaacttctaccccgtatttgccgtgccagaggtgcttgccacctatcacatctttttccaaaactgcaagatacccctatcctgccgtg ccaaccgcagccgagcggacaagcagctggccttgcggcagggcgctgtcatacctgatatcgcctcgctcgacgaagtgccaaaa atctttgagggtcttggacgcgacgagaagcgcgcggcaaacgctctgcaacaagaaaacagcgaaaatgaaagtcactgtggagt gctggtggaacttgagggtgacaacgcgcgcctagccgtgctgaaacgcagcatcgaggtcacccactttgcctacccggcacttaa cctaccccccaaggttatgagcacagtcatgagcgagctgatcgtgcgccgtgcacgacccctggagagggatgcaaacttgcaag aacaaaccgaggagggcctacccgcagttggcgatgagcagctggcgcgctggcttgagacgcgcgagcctgccgacttggagg agcgacgcaagctaatgatggccgcagtgcttgttaccgtggagcttgagtgcatgcagcggttctttgctgacccggagatgcagcg caagctagaggaaacgttgcactacacctttcgccagggctacgtgcgccaggcctgcaaaatttccaacgtggagctctgcaacctg gtctcctaccttggaattttgcacgaaaaccgccttgggcaaaacgtgcttcattccacgctcaagggcgaggcgcgccgcgactacg tccgcgactgcgtttacttatttctgtgctacacctggcaaacggccatgggcgtgtggcagcagtgcctggaggagcgcaacctgaa ggagctgcagaagctgctaaagcaaaacttgaaggacctatggacggccttcaacgagcgctccgtggccgcgcacctggcggac attatcttccccgaacgcctgcttaaaaccctgcaacagggtctgccagacttcaccagtcaaagcatgttgcaaaactttaggaacttta tcctagagcgttcaggaattctgcccgccacctgctgtgcgcttcctagcgactttgtgcccattaagtaccgtgaatgccctccgccgct ttggggtcactgctaccttntgcagctagccaactaccttgcctaccactccgacatcatggaagacgtgagcggtgacggcctactgg agtgtcactgtcgctgcaacctatgcaccccgcaccgctccctggtctgcaattcacaactgcttagcgaaagtcaaattatcggtacctt tgagctgcagggtccctcgcctgacgaaaagtccgcggctccggggttgaaactcactccggggctgtggacgtcggcttaccttcg caaatttgtacctgaggactaccacgcccacgagattaggttctacgaagaccaatcccgcccgccaaatgcggagcttaccgcctgc gtcattacccagggccacatccttggccaattgcaagccattaacaaagcccgccaagagtttctgctacgaaagggacggggggttt acttggacccccagtccggcgaggagctcaacccaatccccccgccgccgcagccctatcagcagccgcgggcccttgcttcccag gatggcacccaaaaagaagctgcagctgccgccgccgccacccacggacgaggaggaatactgggacagtcaggcagaggagg ttggacgaggaggaggagatgatggaagactgggacagcctagacgaggaagcttccgaggccgaagaggtgtcagacgaaac accgtcaccctcggtcgcattcccctcgccggcgccccagaaatcggcaaccgttcccagcattgctacaacctccgctcctcaggcg ccgccggcactgcccgttcgccgacccaaccgtagatgggacaccactggaaccagggccggtaagtctaagcagccgccgccgt tagcccaagagcaacaacagcgccaaggctaccgctcgtggcgcgtgcacaagaacgccatagttgcttgcttgcaagactgtggg ggcaacatctccttcgcccgccgctttcttctctaccatcacggcgtggccttcccccgtaacatcctgcattactaccgtcatctctacag cccctactgcaccggcggcagcggcagcaacagcagcggccacgcagaagcaaaggcgaccggatagcaagactctgacaaag cccaagaaatccacagcggcggcagcagcaggaggaggagcactgcgtctggcgcccaacgaacccgtatcgacccgcgagctt agaaacaggatttttcccactctgtatgctatatttcaacagagcaggggccaagaacaagagctgaaaataaaaaacaggtctctgcg ctccctcacccgcagctgcctgtatcacaaaagcgaagatcagcttcggcgcacgctggaagacgcggaggctctcttcagcaaata ctgcgcgctgactcttaaggactagtttcgcgccctttctcaaatttaagcgcgaaaactacgtcatctccagcggccacacccggcgc cagcacctgtcgtcagcgccattatgagcaaggaaattcccacgccctacatgtggagttaccagccacaaatgggacttgcggctgg agctgcccaagactactcaacccgaataaactacatgagcgcgggaccccacatgatatcccgggtcaacggaatccgcgcccacc gaaaccgaattctcctcgaacaggcggctattaccaccacacctcgtaataaccttaatccccgtagttggcccgctgccctggtgtacc aggaaagtcccgctcccaccactgtggtacttcccagagacgcccaggccgaagttcagatgactaactcaggggcgcagcttgcg ggcggctttcgtcacagggtgcggtcgcccgggcagggtataactcacctgaaaatcagagggcgaggtattcagctcaacgacga gtcggtgagctcctctcttggtctccgtccggacgggacatttcagatcggcggcgctggccgctcttcatttacgccccgtcaggcga tcctaactctgcagacctcgtcctcggagccgcgctccggaggcattggaactctacaatttattgaggagttcgtgccttcggtttactc aaccccttttctggacctcccggccactacccggaccagtttattcccaactttgacgcggtaaaagactcggcggacggctacgactg acagatctgagctcgcggccgcgatatcgctagcgaagttcctattctctagaaagtataggaacttcgatcctctagagtcgacctgca ggcatgcaagcttggcactgcaataaattacttacttaaaatcagtcagcaaatctttgtccagcttattcagcatcacctcctttccctcctc ccaactctggtatttcagcagccttttagctgcgaactttctccaaagtctaaatgggatgtcaaattcctcatgttcttgtccctccgcaccc actatcttcatattgttgcagatgaaacgcgccagaccgtctgaagacaccttcaaccctgtgtacccatatgacacggaaaccggccct ccaactgtgcctttccttacccctccctttgtgtcgccaaatgggttccaagaaagtccccccggagtgctttctttgcgtctttcagaacct ttggttacctcacacggcatgcttgcgctaaaaatgggcagcggcctgtccctggatcaggcaggcaaccttacatcaaatacaatcac tgtttctcaaccgctaaaaaaaacaaagtccaatataactttggaaacatccgcgccccttacagtcagctcaggcgccctaaccatggc cacaacttcgcctttggtggtctctgacaacactcttaccatgcaatcacaagcaccgctaaccgtgcaagactcaaaacttagcattgct accaaagagccacttacagtgttagatggaaaactggccctgcagacatcagcccccctctctgccactgataacaacgccctcactat cactgcctcacctcctcttactactgcaaatggtagtctggctgttaccatggaaaacccactttacaacaacaatggaaaacttgggctc aaaattggcggtcctttgcaagtggccaccgactcacatgcactaacactaggtactggtcagggggttgcagttcataacaatttgcta catacaaaagttacaggcgcaatagggtttgatacatctggcaacatggaacttaaaactggagatggcctctatgtggatagcgccgg tcctaaccaaaaactacatattaatctaaataccacaaaaggccttgcttttgacaacaccgcaataacaattaacgctggaaaagggttg gaatttgaaacagactcctcaaacggaaatcccataaaaacaaaaattggatcaggcatacaatataataccaatggagctatggttgc aaaacttggaacaggcctcagttttgacagctccggagccataacaatgggcagcataaacaatgacagacttactctttggacaacac cagacccatccccaaattgcagaattgcttcagataaagactgcaagctaactctggcgctaacaaaatgtggcagtcaaattttgggc actgtttcagctttggcagtatcaggtaatatggcctccatcaatggaactctaagcagtgtaaacttggttcttagatttgatgacaacgg agtgcttatgtcaaattcatcactggacaaacagtattggaactttagaaacggggactccactaacggtcaaccatacacttatgctgtt gggtttatgccaaacctaaaagcttacccaaaaactcaaagtaaaactgcaaaaagtaatattgttagccaggtgtatcttaatggtgaca agtctaaaccattgcattttactattacgctaaatggaacagatgaaaccaaccaagtaagcaaatactcaatatcattcagttggtcctgg aacagtggacaatacactaatgacaaatttgccaccaattcctataccttctcctacattgcccaggaataaagaatcgtgaacctgttgc atgttatgtttcaacgtgtttatttttcaattcgtattagtcatcgctattaccatggtgatgcggttttggcagtacatcaatgggcgtggatag cggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaat gtcgtaacaactccgccccattgacgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagagctggtttagtgaaccg tcagatccgctagagatccaccatgtttgtctttctcgtgctgctgcccctcgtgagcagccagtgcgtcaatctgacaacaaggaccca gctgccccccgcctacaccaactccttcacaagaggcgtgtattaccccgataaggtcttcagatccagcgtcctccacagcacccaa gatttgtttctgcctttcttcagcaacgtgacatggttccacgccattcatgtcagcggcacaaacggcacaaagaggtttgacaacccc gtgctccccttcaacgacggcgtgtacttcgccagcacagagaaatccaatatcattaggggctggatcttcggcacaacactggattc caagacccagtctctgctcattgtgaataacgccaccaacgtggtgattaaggtctgtgagtttcagttctgcaacgacccctttctggga gtctactaccacaagaataataagagctggatggagtccgagtttagggtgtacagctccgccaacaactgtaccttcgaatacgtgtc ccagcctttcctcatggatctggagggcaagcaaggcaatttcaaaaatctgagagagttcgtgttcaaaaacattgatggatacttcaa aatctacagcaagcatacccccattaatctggtgagggatctgccccaaggattctccgctctggaacctctggtggatctgcccattgg cattaacatcacaagattccagaccctcctcgccctccatagatcctatctgacccccggcgactcctccagcggatggacagccgga gctgccgcctactacgtgggctatctgcagccaagaacctttctgctgaagtacaacgagaacggcaccatcacagacgctgtcgatt gcgctctcgaccctctgagcgagaccaaatgcacactgaagagcttcaccgtggaaaagggcatctatcagaccagcaacttcagag tgcagcctaccgagagcattgtgaggtttcccaacatcaccaatctgtgtcctttcggcgaggtctttaatgccacaaggttcgcttccgt gtatgcttggaataggaagaggatcagcaattgcgtcgccgactattccgtcctctataacagcgcctccttctccaccttcaaatgttatg gcgtgtcccccaccaagctcaacgacctctgcttcaccaatgtgtacgctgactccttcgtcattaggggcgacgaggtgaggcaaatt gcccccggccagaccggcaagattgctgattacaactacaaactgcccgacgattttaccggctgcgtgatcgcttggaactccaaca atctggactccaaagtgggcggaaactacaattacctctacagactctttagaaaaagcaatctgaagcccttcgagagagacatctcc accgaaatctaccaagccggaagcacaccttgcaatggcgtcgagggatttaactgctacttccctctgcagagctacggctttcaacc taccaacggcgtcggatatcaaccctatagggtggtcgtgctgagctttgaactgctgcatgctcccgccaccgtctgcggacctaaga agagcaccaatctcgtcaaaaacaagtgcgtgaacttcaacttcaatggactgaccggcaccggcgtgctgaccgagagcaataaga agtttctgcccttccagcagttcggaagggatattgccgataccacagatgctgtgagggacccccaaaccctcgagattctggatatc accccttgcagcttcggaggagtgtccgtgatcacccccggaacaaacacctccaatcaagtggctgtgctgtaccaagacgtgaact gcacagaagtccccgtggccatccatgccgaccagctgacccctacatggagagtgtactccaccggcagcaatgtgttccagacaa gagccggatgcctcattggagctgaacacgtcaacaacagctacgagtgcgacattcccatcggcgccggcatttgtgcctcctatca gacccagaccaacagcccaagaagggctagaagcgtcgcttcccaatccatcattgcctacaccatgtctctgggagccgaaaactc cgtcgcctactccaacaatagcatcgccatccccaccaattttaccatctccgtgaccacagagattctgcccgtgtccatgacaaagac atccgtggactgcaccatgtacatctgtggcgacagcaccgagtgtagcaatctgctgctgcaatatggcagcttctgcacccagctga acagagccctcaccggcatcgccgtcgaacaagacaagaacacccaagaggtgttcgcccaagtgaagcaaatctacaagacccc ccctatcaaagatttcggaggattcaactttagccagattctgcccgatcctagcaagccttccaagaggagcttcatcgaggatctgct gtttaataaggtgacactggccgacgctggcttcattaaacagtacggcgattgtctgggcgacatcgctgctagggatctgatctgcg ctcagaagttcaacggactgacagtcctccctcctctgctgaccgacgagatgatcgctcagtataccagcgctctgctggctggaacc attaccagcggctggacattcggcgctggagccgccctccaaattccctttgccatgcagatggcctatagattcaacggcattggcgt cacccaaaatgtgctgtatgaaaatcagaagctgattgctaaccaattcaatagcgccattggcaagatccaagactctctgagctccac agccagcgccctcggaaagctgcaagacgtggtgaatcaaaacgcccaagctctgaacacactggtgaaacagctcagcagcaact ttggagccatcagcagcgtgctcaatgatatcctctctaggctggacaaagtggaggccgaagtccagatcgatagactcatcaccgg cagactccaatctctgcagacatacgtcacccaacagctcattagagctgccgaaatcagagcctccgccaatctggccgccaccaa gatgtccgagtgcgtgctgggacagagcaagagagtggacttctgtggcaagggataccatctgatgagcttcccccagagcgctcc ccatggagtggtctttctgcatgtcacatacgtgcccgcccaagagaagaacttcaccaccgctcccgccatttgccacgatggaaag gcccactttcccagagaaggagtgttcgtgagcaacggcacacactggtttgtcacccagagaaatttttacgagccccagattatcac caccgacaacaccttcgtgtccggaaactgcgatgtcgtgattggcatcgtgaacaacacagtctacgaccctctgcagcccgaactc gacagcttcaaggaagagctggacaagtacttcaagaatcacacatcccccgacgtggatctgggcgacattagcggcattaatgcct ccgtcgtcaacattcagaaggagattgatagactgaatgaagtcgccaagaacctcaatgagtctctgattgatctgcaagagctgggc aagtacgagcaatacatcaaatggccttggtacatctggctgggattcatcgctggactcatcgccatcgtgatggtcaccattatgctgt gttgcatgaccagctgctgcagctgtctgaagggctgctgcagctgcggaagctgctgcaagtttgacgaagacgactccgagcccg tgctgaagggcgtcaagctgcattatacataaactagtgctggaattcgcccttatagagtgctggaattcgcccttatagagtgctggaa ttcgcccttatatctagtaacggccgccagtgtgctggaattcgcccttataacttcgtatagcatacattatacgaagttattgttgacaatt aatcatcggcatagtatatcggcatagtataatacgacaaggtgaggaactaaaccatggccaagttgaccagtgccgttccggtgctc accgcgcgcgacgtcgccggagcggtcgagttctggaccgaccggctcgggttctcccgggacttcgtggaggacgacttcgccg gtgtggtccgggacgacgtgaccctgttcatcagcgcggtccaggaccaggtggtgccggacaacaccctggcctgggtgtgggtg cgcggcctggacgagctgtacgccgagtggtcggaggtcgtgtccacgaacttccgggacgcctccgggccggccatgaccgaga tcggcgagcagccgtgggggcgggagttcgccctgcgcgacccggccggcaactgcgtgcacttcgtggccgaggagcaggact gaataacttcgtatagcatacattatacgaagttataagggcgaattctgcagatatccatcctttaaaaaacctcccacacctccccctga acctgaaacataaaatgaatgcaattgttgttgttaacttgtttattgcagcttataatggttacaaataaagcaatagcatcacaaatttcac aaataaagcatttttttcactgcattctagttgtggtttgtccaaactcatcaatgtatcttaacaacgtgtttatttttcaattgcagaaagaatt gcagaaaatttcaagtcatttttcattcagtagtatagccccaccaccacatagcttatactaatcaccgtaccttaatcaaactcacagaa ccctagtattcaacctgccacctccctcccaacacacagagtacacagtcctttctccccggctggccttaaacagcatcatatcatggg taacagacatattcttaggtgttatattccacacggtctcctgtcgagccaaacgctcatcagtgatgttaataaactccccgggcagctc gcttaagttcatgtcgctgtccagctgctgagccacaggctgctgtccaacttgcggttgctcaacgggcggcgaaggagaagtccac gcctacatgggggtagagtcataatcgtgcatcaggatagggcggtggtgctgcagcagcgcgcgaataaactgctgccgccgccg ctccgtcctgcaggaatacaacatggcagtggtctcctcagcgatgattcgcaccgcccgcagcataaggcgccttgtcctccgggca cagcagcgcaccctgatctcacttaagtcagcacagtaactgcagcacagtaccacaatattgtttaaaatcccacagtgcaaggcgct gtatccaaagctcatggcggggaccacagaacccacgtggccatcataccacaagcgcaggtagattaagtggcgacccctcataa acacgctggacataaacattacctcttttggcatgttgtaattcaccacctcccggtaccatataaacctctgattaaacatggcgccatcc accaccatcctaaaccagctggccaaaacctgcccgccggctatgcactgcagggaaccgggactggaacaatgacagtggagag cccaggactcgtaaccatggatcatcatgctcgtcatgatatcaatgttggcacaacacaggcacacgtgcatacacttcctcaggatta caagctcctcccgcgtcagaaccatatcccagggaacaacccattcctgaatcagcgtaaatcccacactgcagggaagacctcgca cgtaactcacgttgtgcattgtcaaagtgttacattcgggcagcagcggatgatcctccagtatggtagcgcgtgtctctgtctcaaaag gaggtaggcgatccctactgtacggagtgcgccgagacaaccgagatcgtgttggtcgtagtgtcatgccaaatggaacgccggac gtagtcatatttcctgaagcaaaaccaggtgcgggcgtgacaaacagatctgcgtctccggtctcgtcgcttagctcgctctgtgtagta gttgtagtatatccactctctcaaagcatccaggcgccccctggcttcgggttctatgtaaactccttcatgcgccgctgccctgataacat ccaccaccgcagaataagccacacccagccaacctacacattcgttctgcgagtcacacacgggaggagcgggaagagctggaag aaccatgttttttttttttattccaaaagattatccaaaacctcaaaatgaagatctattaagtgaacgcgctcccctccggtggcgtggtcaa actctacagccaaagaacagataatggcatttgtaagatgttgcacaatggcttccaaaaggcaaactgccctcacgtccaagtggacg taaaggctaaacccttcanggtgaatctcctctataaacattccagcaccttcaaccatgcccaaataattttcatctcgccaccttatcaat atgtctctaagcaaatcccgaatattaagtccggccattgtaaaaatctgctccagagcgccctccaccttcagcctcaagcagcgaatc atgattgcaaaaattcaggttcctcacagacctgtataagattcaaaagcggaacattaacaaaaataccgcgatcccgtaggtcccttc gcagggccagctgaacataatcgtgcaggtctgcacggaccagcgcggccacttccccgccaggaaccatgacaaaagaacccac actgattatgacacgcatactcggagctatgctaaccagcgtagccccgatgtaagcttgttgcatgggcggcgatataaaatgcaagg tactgctcaaaaaatcaggcaaagcctcgcgcaaaaaagcaagcacatcgtagtcatgctcatgcagataaaggcaggtaagttccg gaaccaccacagaaaaagacaccatttttctctcaaacatgtctgcgggttcctgcataaacacaaaataaaataacaaaaaaaaaaaa acatttaaacattagaagcctgtcttacaacaggaaaaacaacccttataagcataagacggactacggccatgccggcgtgaccgta aaaaaactggtcaccgtgattaaaaagcaccaccgacagttcctcggtcatgtccggagtcataatgtaagactcggtaaacacatcag gttggttaacatcggtcagtgctaaaaagcgaccgaaatagcccgggggaatacatacccgcaggcgtagagacaacattacagccc ccataggaggtataacaaaattaataggagagaaaaacacataaacccctgaaaaaccctcctgcccctaggcaaaatagcaccctc ccgctccagaacaacatacagcgcttccacagcggcagccataacagtcagccttaccagtaaaaaaacctattaaaaaacaccactc gacacggcaccagctcaatcagtcacagtgtaaaaagggccaagtacagagcgagtatatataggactaaaaaatgacgtaacggtt aaagtccacaaaaaccacccagaaaaccgcacgcgaacctacgcccagaaacgaaagccaaaaaacccacaacttcctcaaatctt cacttccgttttcccacgatacgtcacttcccattttaaaaaaaaactacaattcccaatacatgcaagttactccgccctaaaacctacgtc acccgccccgttcccacgccccgcgccacgtcacaaactccaccccctcattatcatattggcttcaatccaaaataaggtatattattga tgatggcgat

[0493] SEQ ID NO:27 nucleic acid sequence that can encode a SC-Ad-TcdA / B virus cgccatcatcaataatataccttattttggattgaagccaatatgataatgagggggtggagtttgtgacgtggcgcggggcgtgggaac ggggcgggtgacgtagtagtgtggcggaagtgtgatgttgtaagtgtggcggaacacatgtaagcgccggatgtggtaaaagtgacg tttttggtgtgcgccggtgtacacgggaagtgacaattttcgcgcggttttaggcggatgttgtagtaaatttgggcgtaaccaagtaatat ttggccattttcgcgggaaaactgaataagaggaagtgaaatctgaataattctgtgttactcatagcgcgtaatatttgtctagggccgc ggggactttgaccgtttacgtggagactcgcccaggtgtttttctcaggtgttttccgcgttccgggtcaaagttggcgttttattattatagt cagctgacgcgcagtgtatttatacccggtgagttcctcaagaggccactcttgagtgccagcgagtagagttttctcctccgagccgct ccgacaccgggactgaaaatgagacatattatctgccacggaggtgttattaccgaagaaatggccgccagtcttttggaccagctgat cgaagaggtactggctgataatcttccacctcctagccattttgaaccacctacccttcacgaactgtatgatttagacgtgacggccccc gaagatcccaacgaggaggcggtttcgcagatttttcccgagtctgtaatgttggcggtgcaggaagggattgacttattcacttttccgc cggcgcccggttctccggagccgcctcacctttcccggcagcccgagcagccggagcagagagccttgggtccggtttctatgcca aaccttgtgccggaggtgatcgatcttacctgccacgaggctggctttccacccagtgacgacgaggatgaagagggtgaggagtttg tgttagattatgtggagcaccccgggcacggttgcaggtcttgtcattatcaccggaggaatacgggggacccagatattatgtgttcgc tttgctatatgaggacctgtggcatgtttgtctacagtaagtgaaaaattatgggcagtgggtgatagagtggtgggtttggtgtggtaatt tttttttaatttttacagttttgtggtttaaagaattttgtattgtgattttttaaaaggtcctgtgtctgaacctgagcctgagcccgagccagaa ccggagcctgcaagacctacccggcgtcctaaattggtgcctgctatcctgagacgcccgacatcacctgtgtctagagaatgcaata gtagtacggatagctgtgactccggtccttctaacacacctcctgagatacacccggtggtcccgctgtgccccattaaaccagttgcc gtgagagttggtgggcgtcgccaggctgtggaatgtatcgaggacttgcttaacgagtctgggcaacctttggacttgagctgtaaacg ccccaggccataaggtgtaaacctgtgattgcgtgtgtggttaacgcctttgtttgctgaatgagttgatgtaagtttaataaagggtgaga taatgtttaacttgcatggcgtgttaaatggggcggggcttaaagggtatataatgcgccgtgggctaatcttggttacatctgacctcatg gaggcttgggagtgtttggaagatttttctgctgtgcgtaacttgctggaacagagctctaacagtacctcttggttttggaggtttctgtgg ggctcctcccaggcaaagttagtctgcagaattaaggaggattacaagtgggaatttgaagagcttttgaaatcctgtggtgagctgtttg attctttgaatctgggtcaccaggcgcttttccaagagaaggtcatcaagactttggatttttccacaccggggcgcgctgcggctgctgt tgcttttttgagttttataaaggataaatggagcgaagaaacccatctgagcggggggtacctgctggattttctggccatgcatctgtgg agagcggtggtgagacacaagaatcgcctgctactgttgtcttccgtccgcccggcaataataccgacggaggagcaacagcagga ggaagccaggcggcggcggcggcaggagcagagcccatggaacccgagagccggcctggaccctcgggaatgaatgttgtaca ggtggctgaactgtttccagaactgagacgcattttaaccattaacgaggatgggcaggggctaaagggggtaaagagggagcggg gggcttctgaggctacagaggaggctaggaatctaacttttagcttaatgaccagacaccgtcctgagtgtgttacttttcagcagattaa ggataattgcgctaatgagcttgatctgctggcgcagaagtattccataaagcagctgaccacttactggctgcagccaggggatgattt tgaggaggctattagggtatatgcaaaggtggcacttaggccagattgcaagtacaagattagcaaacttgtaaatatcaggaattgttg ctacatttctgggaacggggccgaggtggagatagatacggaggatagggtggcctttagatgtagcatgataaatatgtggccggg ggtgcttggcatggacggggtggttattatgaatgtgaggtttactggtcccaattttagcggtacggttttcctggccaataccaatcttat cctacacggtgtaagcttctatgggtttaacaatacctgtgtggaagcctggaccgatgtaagggttcggggctgtgccttttactgctgc tggaagggggtggtgtgtcgccccaaaagcagggcttcaattaagaaatgcctgtttgaaaggtgtaccttgggtatcctgtctgaggg taactccagggtgcgccacaatgtggcctccgactgtggttgctttatgctagtgaaaagcgtggctgtgattaagcataacatggtgtgt ggcaactgcgaggacagggcctctcagatgctgacctgctcggacggcaactgtcacttgctgaagaccattcacgtagccagccac tctcgcaaggcctggccagtgtttgagcacaacatactgacccgctgttccttgcatttgggtaacaggaggggggtgttcctaccttac caatgcaatttgagtcacactaagatattgcttgagcccgagagcatgtccaaggtgaacctgaacggggtgtttgacatgaccatgaa gatctggaaggtgctgaggtacgatgagacccgcaccaggtgcagaccctgcgagtgtggcggtaaacatattaggaaccagcctgt gatgctggatgtgaccgaggagctgaggcccgatcacttggtgctggcctgcacccgcgctgagtttggctctagcgatgaagataca gattgaggtactgaaatgtgtgggcgtggcttaagggtgggaaagaatatataaggtgggggtctcatgtagttttgtatctgttttgcag cagccgccgccatgagcgccaactcgtttgatggaagcattgtgagctcatatttgacaacgcgcatgcccccatgggccggggtgc gtcagaatgtgatgggctccagcattgatggtcgccccgtcctgcccgcaaactctactaccttgacctacgagaccgtgtctggaacg ccgttggagactgcagcctccgccgccgcttcagccgctgcagccaccgcccgcgggattgtgactgactttgctttcctgagcccgc ttgcaagcagtgcagcttcccgttcatccgcccgcgatgacaagttgacggctcttttggcacaattggattctttgacccgggaacttaa tgtcgtttctcagcagctgttggatctgcgccagcaggtttctgccctgaaggcttcctcccctcccaatgcggtttaaaacataaataaaa accagactctgtttggatttggatcaagcaagtgtcttgctgtctttatttaggggttttgcgcgcgcggtaggcccgggaccagcggtct cggtcgttgagggtcctgtgtattttttccaggacgtggtaaaggtgactctggatgttcagatacatgggcataagcccgtctctggggt ggaggtagcaccactgcagagcttcatgctgcggggtggtgttgtagatgatccagtcgtagcaggagcgctgggcgtggtgcctaa aaatgtctttcagtagcaagctgattgccaggggcaggcccttggtgtaagtgtttacaaagcggttaagctgggatgggtgcatacgt ggggatatgagatgcatcttggactgtatttttaggttggctatgttcccagccatatccctccggggattcatgttgtgcagaaccaccag cacagtgtatccggtgcacttgggaaatttgtcatgtagcttagaaggaaatgcgtggaagaacttggagacgcccttgtgacctccaa gattttccatgcattcgtccataatgatggcaatgggcccacgggcggcggcctgggcgaagatatttctgggatcactaacgtcatagt tgtgttccaggatgagatcgtcataggccatttttacaaagcgcgggcggagggtgccagactgcggtataatggttccatccggccca ggggcgtagttaccctcacagatttgcatttcccacgctttgagttcagatggggggatcatgtctacctgcggggcgatgaagaaaac cgtttccggggtaggggagatcagctgggaagaaagcaggttcctaagcagctgcgacttaccgcagccggtgggcccgtaaatca cacctattaccggctgcaactggtagttaagagagctgcagctgccgtcatccctgagcaggggggccacttcgttaagcatgtccctg acttgcatgttttccctgaccaaatccgccagaaggcgctcgccgcccagcgatagcagttcttgcaaggaagcaaagtttttcaacgg tttgaggccgtccgccgtaggcatgcttttgagcgtttgaccaagcagttccaggcggtcccacagctcggtcacgtgctctacggcat ctcgatccagcatatctcctcgtttcgcgggttggggcggctttcgctgtacggcagtagtcggtgctcgtccagacgggccagggtca tgtctttccacgggcgcagggtcctcgtcagcgtagtctgggtcacggtgaaggggtgcgctccgggttgcgcgctggccagggtgc gcttgaggctggtcctgctggtgctgaagcgctgccggtcttcgccctgcgcgtcggccaggtagcatttgaccatggtgtcatagtcc agcccctccgcggcgtggcccttggcgcgcagcttgcccttggaggaggcgccgcacgaggggcagtgcagacttttaagggcgt agagcttgggcgcgagaaataccgattccggggagtaggcatccgcgccgcaggccccgcagacggtctcgcattccacgagcca ggtgagctctggccgttcggggtcaaaaaccaggtttcccccatgctttttgatgcgtttcttacctctggtttccatgagccggtgtccac gctcggtgacgaaaaggctgtccgtgtccccgtatacagacttgagaggcctgtcctcgagcggtgttccgcggtcctcctcgtataga aactcggaccactctgagacgaaggctcgcgtccaggccagcacgaaggaggctaagtgggaggggtagcggtcgttgtccacta gggggtccactcgctccagggtgtgaagacacatgtcgccctcttcggcatcaaggaaggtgattggtttataggtgtaggccacgtg accgggtgttcctgaaggggggctataaaagggggtgggggcgcgttcgtcctcactctcttccgcatcgctgtctgcgagggccag ctgttggggtgagtactccctctcaaaagcgggcatgacttctgcgctaagattgtcagtttccaaaaacgaggaggatttgatattcacc tggcccgcggtgatgcctttgagggtggccgcgtccatctggtcagaaaagacaatctttttgttgtcaagcttggtggcaaacgaccc gtagagggcgttggacagcaacttggcgatggagcgcagggtttggtttttgtcgcgatcggcgcgctccttggccgcgatgtttagct gcacgtattcgcgcgcaacgcaccgccattcgggaaagacggtggtgcgctcgtcgggcactaggtgcacgcgccaaccgcggtt gtgcagggtgacaaggtcaacgctggtggctacctctccgcgtaggcgctcgttggtccagcagaggcggccgcccttgcgcgagc agaatggcggtagtgggtctagctgcgtctcgtccggggggtctgcgtccacggtaaagaccccgggcagcaggcgcgcgtcgaa gtagtctatcttgcatccttgcaagtctagcgcctgctgccatgcgcgggcggcaagcgcgcgctcgtatgggttgagtgggggaccc catggcatggggtgggtgagcgcggaggcgtacatgccgcaaatgtcgtaaacgtagaggggctctctgagtattccaagatatgta gggtagcatcttccaccgcggatgctggcgcgcacgtaatcgtatagttcgtgcgagggagcgaggaggtcgggaccgaggttgct acgggcgggctgctctgctcggaagactatctgcctgaagatggcatgtgagttggatgatatggttggacgctggaagacgttgaag ctggcgtctgtgagacctaccgcgtcacgcacgaaggaggcgtaggagtcgcgcagcttgttgaccagctcggcggtgacctgcac gtctagggcgcagtagtccagggtttccttgatgatgtcatacttatcctgtcccttttttttccacagctcgcggttgaggacaaactcttc gcggtctttccagtactcttggatcggaaacccgtcggcctccgaacggtaagagcctancatgtagaactggttgacggcctggtag gcgcagcatcccttttctacgggtagcgcgtatgcctgcgcggccttccggagcgaggtgtgggtgagcgcaaaggtgtccctaacc atgactttgaggtactggtatttgaagtcagtgtcgtcgcatccgccctgctcccagagcaaaaagtccgtgcgctttttggaacgcggg tttggcagggcgaaggtgacatcgttgaagagtatctttcccgcgcgaggcataaagttgcgtgtgatgcggaagggtcccggcacct cggaacggttgttaattacctgggcggcgagcacgatctcgtcaaagccgttgatgttgtggcccacaatgtaaagttccaagaagcg cgggatgcccttgatggaaggcaattttttaagttcctcgtaggtgagctcttcaggggagctgagcccgtgctctgaaagggcccagt ctgcaagatgagggttggaagcgacgaatgagctccacaggtcacgggccattagcatttgcaggtggtcgcgaaaggtcctaaact ggcgacctatggccattttttctggggtgatgcagtagaaggtaagcgggtcttgttcccagcggtcccatccaaggtccgcggctagg tctcgcgcggcggtcactagaggctcatctccgccgaacttcatgaccagcatgaagggcacgagctgcttcccaaaggcccccatc caagtataggtctctacatcgtaggtgacaaagagacgctcggtgcgaggatgcgagccgatcgggaagaactggatctcccgcca ccagttggaggagtggctgttgatgtggtgaaagtagaagtccctgcgacgggccgaacactcgtgctggcttttgtaaaaacgtgcg cagtactggcagcggtgcacgggctgtacatcctgcacgaggttgacctgacgaccgcgcacaaggaagcagagtgggaatttgag cccctcgcctggcgggtttggctggtggtcttctacttcggctgcttgtccttgaccgtctggctgctcgaggggagttacggtggatcg gaccaccacgccgcgcgagcccaaagtccagatgtccgcgcgcggcggtcggagcttgatgacaacatcgcgcagatgggagct gtccatggtctggagctcccgcggcgtcaggtcaggcgggagctcctgcaggtttacctcgcatagccgggtcagggcgcgggcta ggtccaggtgatacctgatttccaggggctggttggtggcggcgtcgatggcttgcaagaggccgcatccccgcggcgcgactacg gtaccgcgcggcgggcggtgggccgcgggggtgtccttggatgatgcatctaaaagcggtgacgcgggcgggcccccggaggta gggggggctcgggacccgccgggagagggggcaggggcacgtcggcgccgcgcgcgggcaggagctggtgctgcgcgcgg aggttgctggcgaacgcgacgacgcggcggttgatctcctgaatctggcgcctctgcgtgaagacgacgggcccggtgagcttgaa cctgaaagagagttcgacagaatcaatttcggtgtcgttgacggcggcctggcgcaaaatctcctgcacgtctcctgagttgtcttgata ggcgatctcggccatgaactgctcgatctcttcctcctggagatctccgcgtccggctcgctccacggtggcggcgaggtcgttggag atgcgggccatgagctgcgagaaggcgttgaggcctccctcgttccagacgcggctgtagaccacgcccccttcggcatcgcgggc gcgcatgaccacctgcgcgagattgagctccacgtgccgggcgaagacggcgtagtttcgcaggcgctgaaagaggtagttgagg gtggtggcggtgtgttctgccacgaagaagtacataacccagcgccgcaacgtggattcgttgatatcccccaaggcctcaaggcgct ccatggcctcgtagaagtccacggcgaagttgaaaaactgggagttgcgcgccgacacggttaactcctcctccagaagacggatga gctcggcgacagtgtcgcgcacctcgcgctcaaaggctacaggggcctcttcttcttcttcaatctcctcttccataagggcctccccttc ttcttcttctggcggcggtgggggaggggggacacggcggcgacgacggcgcaccgggaggcggtcgacaaagcgctcgatcat ctccccgcggcgacggcgcatggtctcggtgacggcgcggccgttctcgcgggggcgcagttggaagacgccgcccgtcatgtcc cggtatgggtggcggggggctgccgtgcggcagggatacggcgctaacgatgcatctcaacaattgttgtgtaggtactccgccac cgagggacctgagcgagtccgcatcgaccggatcggaaaacctctcgagaaaggcgtctaaccagtcacagtcgcaaggtaggct gagcaccgtggcgggcggcagcgggcggcggtcggggttgtttctggcggaggtgctgctgatgatgtaattaaagtaggcggtctt gagacggcggatggtcgacagaagcaccatgtccttgggtccggcctgctgaatgcgcaggcggtcggccatgccccaggcttcgt tttgacatcggcgcaggtctttgtagtagtcttgcatgagcctttctaccggcacttcttcttctccttcctcttgtcctgcatctcttgcatctat cgctgcggcggcggcggagtttggccgtaggtggcgccctcttcctcccatgcgtgtgaccccgaagcccctcatcggctgaagca gggccaggtcggcgacaacgcgctcggctaatatggcctgctgcacctgcgtgagggtagactggaagtcgtccatgtccacaaag cggtggtatgcgcccgtgttgatggtgtaagtgcagttggccataacggaccagttaacggtctggtgacccggctgcgagagctcgg tgtacctgagacgcgagtaagcccttgagtcaaagacgtagtcgttgcaagtccgcaccaggtactggtatcccaccaaaaagtgcgg cggcggctggcggtagaggggccagcgtagggtggccggggctccgggggcgaggtcttccaacataaggcgatgatatccgta gatgtacctggacatccaggtgatgccggcggcggtggtggaggcgcgcggaaagtcacggacgcggttccagatgttgcgcagc ggcaaaaagtgctccatggtcgggacgctctggccggtcaggcgcgcgcagtcgttgacgctctagaccgtgcaaaaggagagcct gtaagcgggcactcttccgtggtctggtggataaattcgcaagggtatcatggcggacgaccggggttcgaaccccggatccggccg tccgccgtgatccatgcggttaccgcccgcgtgtcgaacccaggtgtgcgacgtcagacaacgggggagcgctccttttggcttcctt ccaggcgcggcggatgctgcgctagcttttttggccactggccgcgcgcggcgtaagcggttaggctggaaagcgaaagcattaagt ggctcgctccctgtagccggagggttattttccaagggttgagtcgcgggacccccggttcgagtctcgggccggccggactgcggc gaacgggggtttgcctccccgtcatgcaagaccccgcttgcaaattcctccggaaacagggacgagccccttttttgcttttcccagatg catccggtgctgcggcagatgcgcccccctcctcagcagcggcaagagcaagagcagcggcagacatgcagggcaccctcccct cctcctaccgcgtcaggaggggcgacatccgcggttgacgcggcagcagatggtgattacgaacccccgcggcgccgggcccgg cactacctggacttggaggagggcgagggcctggcgcggctaggagcgccctctcctgagcggtacccaagggtgcagctgaagc gtgatacgcgtgaggcgtacgtgccgcggcagaacctgtttcgcgaccgcgagggagaggagcccgaggagatgcgggatcgaa agttccacgcagggcgcgagctgcggcatggcctgaatcgcgagcggttgctgcgcgaggaggactttgagcccgacgcgcgaac cgggattagtcccgcgcgcgcacacgtggcggccgccgacctggtaaccgcatacgagcagacggtgaaccaggagattaactttc aaaaaagctttaacaaccacgtgcgtacgcttgtggcgcgcgaggaggtggctataggactgatgcatctgtgggactttgtaagcgc gctggagcaaaacccaaatagcaagccgctcatggcgcagctgttccttatagtgcagcacagcagggacaacgaggcattcaggg atgcgctgctaaacatagtagagcccgagggccgctggctgctcgatttgataaacatcctgcagagcatagtggtgcaggagcgca gcttgagcctggctgacaaggtggccgccatcaactattccatgcttagcctgggcaagttttacgcccgcaagatataccatacccctt acgttcccatagacaaggaggtaaagatcgaggggttctacatgcgcatggcgctgaaggtgcttaccttgagcgacgacctgggcg tttatcgcaacgagcgcatccacaaggccgtgagcgtgagccggcggcgcgagctcagcgaccgcgagctgatgcacagcctgca aagggccctggctggcacgggcagcggcgatagagaggccgagtcctactttgacgcgggcgctgacctgcgctgggccccaag ccgacgcgccctggaggcagctggggccggacctgggctggcggtggcacccgcgcgcgctggcaacgtcggcggcgtggag gaatatgacgaggacgatgagtacgagccagaggacggcgagtactaagcggtgatgtttctgatcagtcgcggccgcgatatcgct agcgaagttcctattctctagaaagtataggaacttcggatcctctagagtcgaaaaaaaaaaagcatgatgcaaaataaaaaactcac caaggccatggcaccgagcgttggttttcttgtattccccttagtatgcggcgcgcggcgatgtatgaggaaggtcctcctccctcctac gagagtgtggtgagcgcggcgccagtggcggcggcgctgggttctcccttcgatgctcccctggacccgccgtttgtgcctccgcgg tacctgcggcctaccggggggagaaacagcatccgttactctgagttggcacccctattcgacaccacccgtgtgtacctggtggaca acaagtcaacggatgtggcatccctgaactaccagaacgaccacagcaactttctgaccacggtcattcaaaacaatgactacagccc gggggaggcaagcacacagaccatcaatcttgacgaccggtcgcactggggcggcgacctgaaaaccatcctgcataccaacatg ccaaatgtgaacgagttcatgtttaccaataagtttaaggcgcgggtgatggtgtcgcgcttgcctactaaggacaatcaggtggagct gaaatacgagtgggtggagttcacgctgcccgagggcaactactccgagaccatgaccatagaccttatgaacaacgcgatcgtgga gcactacttgaaagtgggcagacagaacggggttctggaaagcgacatcggggtaaagtttgacacccgcaacttcagactggggtt tgaccccgtcactggtcttgtcatgcctggggtatatacaaacgaagccttccatccagacatcattttgctgccaggatgcggggtgga cttcacccacagccgcctgagcaacttgttgggcatccgcaagcggcaacccttccaggagggctttaggatcacctacgatgatctg gagggtggtaacattcccgcactgttggatgtggacgcctaccaggcgagcttgaaagatgacaccgaacagggcgggggtggcg caggcggcagcaacagcagtggcagcggcgcggaagagaactccaacgcggcagccgcggcaatgcagccggtggaggacat gaacgatcatgccattcgcggcgacacctttgccacacgggctgaggagaagcgcgctgaggccgaagcagcggccgaagctgc cgcccccgctgcgcaacccgaggtcgagaagcctcagaagaaaccggtgatcaaacccctgacagaggacagcaagaaacgcag ttacaacctaataagcaatgacagcaccttcacccagtaccgcagctggtaccttgcatacaactacggcgaccctcagaccggaatc cgctcatggaccctgctttgcactcctgacgtaacctgcggctcggagcaggtctactggtcgttgccagacatgatgcaagaccccgt gaccttccgctccacgcgccagatcagcaactttccggtggtgggcgccgagctgttgcccgtgcactccaagagcttctacaacgac caggccgtctactcccaactcatccgccagtttacctctctgacccacgtgttcaatcgctttcccgagaaccagattttggcgcgcccg ccagcccccaccatcaccaccgtcagtgaaaacgttcctgctctcacagatcacgggacgctaccgctgcgcaacagcatcggagg agtccagcgagtgaccattactgacgccagacgccgcacctgcccctacgtttacaaggccctgggcatagtctcgccgcgcgtcct atcgagccgcactttttgagcaagcatgtccatccttatatcgcccagcaataacacaggctggggcctgcgcttcccaagcaagatgtt tggcggggccaagaagcgctccgaccaacacccagtgcgcgtgcgcgggcactaccgcgcgccctggggcgcgcacaaacgcg gccgcactgggcgcaccaccgtcgatgacgccatcgacgcggtggtggaggaggcgcgcaactacacgcccacgccgccgcca gtgtccaccgtggacgcggccattcagaccgtggtgcgcggagcccggcgctacgctaaaatgaagagacggcggaggcgcgta gcacgtcgccaccgccgccgacccggcactgccgcccaacgcgcggcggcggccctgcttaaccgcgcacgtcgcaccggccg acgggcggccatgcgagccgctcgaaggctggccgcgggtattgtcactgtgccccccaggtccaggcgacgagcggccgccgc agcagccgcggccattagtgttatgactcagggtcgcaggggcaacgtgtactgggtgcgcgactcggttagcggcctgcgcgtgc ccgtgcgcacccgccccccgcgcaactagattgcaataaaaaactacttagactcgtactgttgtatgtatccagcggcggcggcgcg catcgaagctatgtccaagcgcaaaatcaaagaagagatgctccaggtcatcgcgccggagatctatggccccccgaagaaggaag agcaggattacaagccccgaaagctaaagcgggtcaaaaagaaaaagaaagatgatgatgatgatgaacttgacgacgaggtggaa ctgttgcacgcgaccgcgcccaggcgacgggtacagtggaaaggtcgacgcgtaagacgtgttttgcgacccgg caeca ccgtagt ctttacgcccggtgagcgctccacccgcacctacaagcgcgtgtatgatgaggtgtacggcgacgaggacctgcttgagcaggcca acgagcgcctcggggagtttgcctacggaaagcggcataaggacatgctggcgttgccgctggacgagggcaacccaacacctag cctaaagcccgtgacactgcagcaggtgctgcccgcgcttgcaccgtccgaagaaaagcgcggcctaaagcgcgagtctggtgact tggcacccaccgtgcagctgatggtacccaagcgtcagcgactggaagatgtcttggaaaaaatgaccgtggagcctgggctggag cccgaggtccgcgtgcggccaatcaagcaggtggcaccgggactgggcgtgcagaccgtggacgttcagatacccaccaccagta gcactagtattgccactgccacagagggcatggagacacaaacgtccccggttgcctcggcggtggcagatgccgcggtgcaggc ggccgctgcggccgcgtccaagacctctacggaggtgcaaacggacccgtggatgtttcgtgtttcagccccccggcgtccgcgcc gttcaaggaagtacggcgccgccagcgcgctactgcccgaatatgccctacatccttccatcgcgcctacccccggctatcgtggcta cacctaccgccccagaagacgagcaactacccgacgccgaaccaccactggaacccgccgccgccgtcgccgtcgccagcccgt gctggccccgatttccgtgcgcagggtggctcgcgaaggaggcaggaccctggtgctgccaacagcgcgctaccaccccagcatc gtttaaaagccggtctttgtggttcttgcagatatggccctcacctgccgcctccgtttcccggtgccgggattccgaggaagaatgcac cgtaggaggggcatggccggccacggcctgacgggcggcatgcgtcgtgcgcaccaccggcggcggcgcgcgtcgcaccgtcg catgcgcgccggtatcctgcccctccttattccactgatcgccgcggcgattggcgccgtgcccggaattgcatccgtggccttgcag gcgcagagacactgattaaaaacaagttacatgtggaaaaatcaaaataaaagtctggactctcacgctcgcttggtcctgtaactatttt gtagaatggaagacatcaactttgcgtcactggccccgcgacacggctcgcgcccgttcatgggaaactggcaagatatcgg caeca gcaatatgagcggtggcgccttcagctggggctcgctgtggagcggcattaaaaatttcggttccgccgttaagaactatggcagcaa agcctggaacagcagcacaggccagatgctgagggacaagttgaaagagcaaaatttccaacaaaaggtggtagatggcctggcct ctggcattagcggggtggtggacctggccaaccaggcagtgcaaaataagattaacagtaagcttgatccccgccctcccgtagagg agcctccaccggccgtggagacagtgtctccagaggggcgtggcgaaaagcgtccgcgacccgacagggaagaaactctggtga cgcaaatagacgagcctccctcgtacgaggaggcactaaagcaaggcctgcccaccacccgtcccatcgcgcccatggctaccgg agtgctgggccagcacacacccgtaacgctggacctgcctccccccgccgacacccagcagaaacctgtgctgccaggcccgtcc gccgttgttgtaacccgtcctagccgcgggtccctgcgccgcgccgccagcggtccgcgatcgttgcggcccgtagccagtggcaa ctggcaaagcacactgaacagcatcgtgggtttgggggtgcaatccctgaagcgccgacgatgcttctgatagctaacgtgtcgtatgt gtgtcatgtatgcgtccatgtcgccgccagaggagctgctgagccgccgcgcgcccgctttccaagatggctaccccttcgatgatgc cgcagtggtcttacatgcacatctcgggccaggacgcctcggagtacctgagccccgggctggtgcagttcgcccgcgccaccgag acgtacttcagcctgaataacaagtttagaaaccccacggtggcgcctacgcacgacgtgaccacagaccggtctcagcgtttgacgc tgcggttcatccccgtggaccgcgaggatactgcgtactcgtacaaggcgcggttcaccctagctgtgggtgataaccgtgtgctaga catggcttccacgtactttgacatccgcggcgtgctggacaggggccctacttttaagccctactctggcactgcctacaacgcactgg cccccaagggtgcccccaactcgtgcgagtgggaacaaaatgaaactgcacaagtggatgctcaagaacttgacgaagaggagaat gaagccaatgaagctcaggcgcgagaacaggaacaagctaagaaaacccatgtatatgcccaggctccactgtccggaataaaaat aactaaagaaggtctacaaataggaactgccgacgccacagtagcaggtgccggcaaagaaattttcgcagacaaaacttttcaacct gaaccacaagtaggagaatctcaatggaacgaagcggatgccacagcagctggtggaagggttcttaaaaagacaactcccatgaa accctgctatggctcatacgctagacccaccaattccaacggcggacagggcgttatggttgaacaaaatggtaaattggaaagtcaa gtcgaaatgcaatttttttccacatccacaaatgccacaaatgaagttaacaatatacaaccaacagttgtattgtacagcgaagatgtaaa catggaaactccagatactcatctttcttataaacctaaaatgggggataaaaatgccaaagtcatgcttggacaacaagcaatgccaaa cagaccaaattacattgcttttagagacaattttattggtctcatgtattacaacagcacaggtaacatgggtgtccttgctggtcaggcatc gcagttgaacgctgttgtagatttgcaagacagaaacacagagctgtcctaccagcttttgcttgattcaattggcgacagaacaagata cttttcaatgtggaatcaagctgttgacagctatgatccagatgtcagaattattgagaaccatggaactgaggatgagttgccaaattatt gctttcctcttggtggaattgggattactgacacttttcaagctgttaaaacaactgctgctaacggggaccaaggcaatactacctggca aaaagattcaacatttgcagaacgcaatgaaataggggtgggaaataactttgccatggaaattaacctgaatgccaacctatggagaa atttcctttactccaatattgcgctgtacctgccagacaagctaaaatacaaccccaccaatgtggaaatatctgacaaccccaacaccta cgactacatgaacaagcgagtggtggctcctgggcttgtagactgctacattaaccttggggcgcgctggtctctggactacatggaca acgttaatccctttaaccacccccgccatgcgggcctgcgttaccgctccatgttgttgggaaacggccgctacgtgccctttcacattc aggtgccccaaaagttttttgccattaaaaacctcctcctcctgccaggctcatacacatatgaatggaacttcaggaaggatgttaacat ggttctgcagagctctctgggaaacgaccttagagttgacggggctagcattaagtttgacagcatttgtctttacgccaccttcttcccca tggcccacaacacggcctccacgctggaagccatgctcagaaatgacaccaacgaccagtcctttaatgactacctttccgccgccaa catgctatatcccatacccgccaacgccaccaacgtgcccatctccatcccatcgcgcaactgggcagcatttcgcggttgggccttca cacgcttgaagacaaaggaaaccccttccctgggatcaggctacgacccttactacacctactctggctccataccataccttgacgga accttctatcttaatcacacctttaagaaggtggccattacttttgactcttctgttagctggccgggcaacgaccgcctgcttactcccaat gagtttgagattaagcgctcagttgacggggagggctataacgtagctcagtgcaacatgacaaaggactggttcctagtgcagatgtt ggccaactacaatattggctaccagggcttctacattccagaaagctacaaagaccgcatgtactcgttcttcagaaacttccagcccat gagccggcaagtggtggacgatactaaatacaaagattatcagcaggttggaattatccaccagcataacaactcaggcttcgtaggct acctcgctcccaccatgcgcgagggacaagcttaccccgctaatgttccctacccactaataggcaaaaccgcggttgatagtattacc cagaaaaagtttctttgcgaccgcaccctgtggcgcatccccttctccagtaactttatgtccatgggtgcgctcacagacctgggccaa aaccttctctacgcaaactccgcccacgcgctagacatgacctttgaggtggatcccatggacgagcccacccttctttatgttttgtttga agtctttgacgtggtccgtgtgcaccagccgcaccgcggcgtcatcgagaccgtgtacctgcgcacgcccttctcggccggcaacgc cacaacataaagaagcaagcaacatcaacaacagctgccgccatgggctccagtgagcaggaactgaaagccattgtcaaagatctt ggttgtgggccatattttttgggcacctatgacaagcgcttcccaggctttgtttccccacacaagctcgcctgcgccatagttaacacgg ccggtcgcgagactgggggcgtacactggatggcctttgcctggaacccgcgctcaaaaacatgctacctctttgagccctttggctttt ctgaccaacgtctcaagcaggtttaccagtttgagtacgagtcactcctgcgccgtagcgccattgcctcttcccccgaccgctgtataa cgctggaaaagtccacccaaagcgtgcaggggcccaactcggccgcctgtggcctattctgctgcatgtttctccacgcctttgccaac tggccccaaactcccatggatcacaaccccaccatgaaccttattaccggggtacccaactccatgcttaacagtccccaggtacagc ccaccctgcgccgcaaccaggaacagctctacagcttcctggagcgccactcgccctacttccgcagccacagtgcgcaaattagga gcgccacttctttttgtcacttgaaaaacatgtaaaaataatgtactaggagacactttcaataaaggcaaatgtttttatttgtacactctcg ggtgattatttacccccacccttgccgtctgcgccgtttaaaaatcaaaggggttctgccgcgcatcgctatgcgccactggcagggac acgttgcgatactggtgtttagtgctccacttaaactcaggcacaaccatccgcggcagctcggtgaagttttcactccacaggctgcgc accatcaccaacgcgtttagcaggtcgggcgccgatatcttgaagtcgcagttggggcctccgccctgcgcgcgcgagttgcgatac acagggttacagcactggaacactatcagcgccgggtggtgcacgctggccagcacgctcttgtcggagatcanatccgcgtccag gtcctccgcgttgctcagggcgaacggagtcaactttggtagctgccttcccaaaaagggtgcatgcccaggctttgagttgcactcgc accgtagtggcatcagaaggtgaccgtgcccagtctgggcgttaggatacagcgcctgcatgaaagccttgatctgcttaaaagccac ctgagcctttgcgccttcagagaagaacatgccgcaagacttgccggaaaactgattggccggacaggccgcgtcatgcacgcagc accttgcgtcggtgttggagatctgcaccacatttcggccccaccggttcttcacgatcttggccttgctagactgctccttcagcgcgcg ctgcccgttttcgctcgtcacatccatttcaatcacgtgctccttatttatcataatgctcccgtgtagacacttaagctcgccttcgatctca gcgcagcggtgcagccacaacgcgcagcccgtgggctcgtggtgcttgtaggttacctctgcaaacgactgcaggtacgcctgcag gaatcgccccatcatcgtcacaaaggtcttgttgctggtgaaggtcagctgcaacccgcggtgctcctcgtttagccaggtcttgcatac ggccgccagagcttccacttggtcaggcagtagcttgaagtttgcctttagatcgttatccacgtggtacttgtccatcaacgcgcgcgc agcctccatgcccttctcccacgcagacacgatcggcaggctcagcgggtttatcaccgtgctttcactttccgcttcactggactcttcc ttttcctcttgcatccgcataccccgcgccactgggtcgtcttcattcagccgccgcaccgtgcgcttacctcccttgccgtgcttgattag caccggtgggttgctgaaacccaccatttgtagcgccacatcttctctttcttcctcgctgtccacgatcacctctggggatggcgggcg ctcgggcttgggagaggggcgcttctttttctttttggacgcaatggccaaatccgccgtcgaggtcgatggccgcgggctgggtgtgc gcggcaccagcgcatcttgtgacgagtcttcttcgtcctcggactcgagacgccgcctcagccgcttttttgggggcgcgcggggag gcggcggcgacggcgacggggacgagacgtcctccatggttggtggacgtcgcgccgcaccgcgtccgcgctcgggggtggttt cgcgctgctcctcttcccgactggccatttccttctcctataggcagaaaaagatcatggagtcagtcgagaaggaggacagcctaacc gccccctttgagttcgccaccaccgcctccaccgatgccgccaacgcgcctaccaccttccccgtcgaggcacccccgcttgaggag gaggaagtgattatcgagcaggacccaggttttgtaagcgaagacgacgaagatcgctcagtaccaacagaggataaaaagcaaga ccaggacgacgcagaggcaaacgaggaacaagtcgggcggggggaccaaaggcatggcgactacctagatgtgggagacgac gtgctgttgaagcatctgcagcgccagtgcgccattatctgcgacgcgttgcaagagcgcagcgatgtgcccctcgccatagcggat gtcagccttgcctacgaacgccacctgttctcaccgcgcgtaccccccaaacgccaagaaaacggcacatgcgagcccaacccgcg cctcaacttctaccccgtatttgccgtgccagaggtgcttgccacctatcacatctttttccaaaactgcaagatacccctatcctgccgtg ccaaccgcagccgagcggacaagcagctggccttgcggcagggcgctgtcatacctgatatcgcctcgctcgacgaagtgccaaaa atctttgagggtcttggacgcgacgagaagcgcgcggcaaacgctctgcaacaagaaaacagcgaaaatgaaagtcactgtggagt gctggtggaacttgagggtgacaacgcgcgcctagccgtgctgaaacgcagcatcgaggtcacccactttgcctacccggcacttaa cctaccccccaaggttatgagcacagtcatgagcgagctgatcgtgcgccgtgcacgacccctggagagggatgcaaacttgcaag aacaaaccgaggagggcctacccgcagttggcgatgagcagctggcgcgctggcttgagacgcgcgagcctgccgacttggagg agcgacgcaagctaatgatggccgcagtgcttgttaccgtggagcttgagtgcatgcagcggttctttgctgacccggagatgcagcg caagctagaggaaacgttgcactacacctttcgccagggctacgtgcgccaggcctgcaaaatttccaacgtggagctctgcaacctg gtctcctaccttggaattttgcacgaaaaccgccttgggcaaaacgtgcttcattccacgctcaagggcgaggcgcgccgcgactacg tccgcgactgcgtttacttatttctgtgctacacctggcaaacggccatgggcgtgtggcagcagtgcctggaggagcgcaacctgaa ggagctgcagaagctgctaaagcaaaacttgaaggacctatggacggccttcaacgagcgctccgtggccgcgcacctggcggac attatcttccccgaacgcctgcttaaaaccctgcaacagggtctgccagacttcaccagtcaaagcatgttgcaaaactttaggaacttta tcctagagcgttcaggaattctgcccgccacctgctgtgcgcttcctagcgactttgtgcccattaagtaccgtgaatgccctccgccgct ttggggtcactgctaccttntgcagctagccaactaccttgcctaccactccgacatcatggaagacgtgagcggtgacggcctactgg agtgtcactgtcgctgcaacctatgcaccccgcaccgctccctggtctgcaattcacaactgcttagcgaaagtcaaattatcggtacctt tgagctgcagggtccctcgcctgacgaaaagtccgcggctccggggttgaaactcactccggggctgtggacgtcggcttaccttcg caaatttgtacctgaggactaccacgcccacgagattaggttctacgaagaccaatcccgcccgccaaatgcggagcttaccgcctgc gtcattacccagggccacatccttggccaattgcaagccattaacaaagcccgccaagagtttctgctacgaaagggacggggggttt acttggacccccagtccggcgaggagctcaacccaatccccccgccgccgcagccctatcagcagccgcgggcccttgcttcccag gatggcacccaaaaagaagctgcagctgccgccgccgccacccacggacgaggaggaatactgggacagtcaggcagaggagg ttttggacgaggaggaggagatgatggaagactgggacagcctagacgaggaagcttccgaggccgaagaggtgtcagacgaaac accgtcaccctcggtcgcattcccctcgccggcgccccagaaatcggcaaccgttcccagcattgctacaacctccgctcctcaggcg ccgccggcactgcccgttcgccgacccaaccgtagatgggacaccactggaaccagggccggtaagtctaagcagccgccgccgt tagcccaagagcaacaacagcgccaaggctaccgctcgtggcgcgtgcacaagaacgccatagttgcttgcttgcaagactgtggg ggcaacatctccttcgcccgccgctttcttctctaccatcacggcgtggccttcccccgtaacatcctgcattactaccgtcatctctacag cccctactgcaccggcggcagcggcagcaacagcagcggccacgcagaagcaaaggcgaccggatagcaagactctgacaaag cccaagaaatccacagcggcggcagcagcaggaggaggagcactgcgtctggcgcccaacgaacccgtatcgacccgcgagctt agaaacaggatttttcccactctgtatgctatatttcaacagagcaggggccaagaacaagagctgaaaataaaaaacaggtctctgcg ctccctcacccgcagctgcctgtatcacaaaagcgaagatcagcttcggcgcacgctggaagacgcggaggctctcttcagcaaata ctgcgcgctgactcttaaggactagtttcgcgccctttctcaaatttaagcgcgaaaactacgtcatctccagcggccacacccggcgc cagcacctgtcgtcagcgccattatgagcaaggaaattcccacgccctacatgtggagttaccagccacaaatgggacttgcggctgg agctgcccaagactactcaacccgaataaactacatgagcgcgggaccccacatgatatcccgggtcaacggaatccgcgcccacc gaaaccgaattctcctcgaacaggcggctattaccaccacacctcgtaataaccttaatccccgtagttggcccgctgccctggtgtacc aggaaagtcccgctcccaccactgtggtacttcccagagacgcccaggccgaagttcagatgactaactcaggggcgcagcttgcg ggcggctttcgtcacagggtgcggtcgcccgggcagggtataactcacctgaaaatcagagggcgaggtattcagctcaacgacga gtcggtgagctcctctcttggtctccgtccggacgggacatttcagatcggcggcgctggccgctcttcatttacgccccgtcaggcga tcctaactctgcagacctcgtcctcggagccgcgctccggaggcattggaactctacaatttattgaggagttcgtgccttcggtttactc aaccccttttctggacctcccggccactacccggaccagtttattcccaactttgacgcggtaaaagactcggcggacggctacgactg acagatctgagctcgcggccgcgatatcgctagcgaagttcctattctctagaaagtataggaacttcgatcctctagagtcgacctgca ggcatgcaagcttggcactgcaataaattacttacttaaaatcagtcagcaaatctttgtccagcttattcagcatcacctcctttccctcctc ccaactctggtatttcagcagccttttagctgcgaactttctccaaagtctaaatgggatgtcaaattcctcatgttcttgtccctccgcaccc actatcttcatattgttgcagatgaaacgcgccagaccgtctgaagacaccttcaaccctgtgtacccatatgacacggaaaccggccct ccaactgtgcctttccttacccctccctttgtgtcgccaaatgggttccaagaaagtccccccggagtgctttctttgcgtctttcagaacct ttggttacctcacacggcatgcttgcgctaaaaatgggcagcggcctgtccctggatcaggcaggcaaccttacatcaaatacaatcac tgtttctcaaccgctaaaaaaaacaaagtccaatataactttggaaacatccgcgccccttacagtcagctcaggcgccctaaccatggc cacaacttcgcctttggtggtctctgacaacactcttaccatgcaatcacaagcaccgctaaccgtgcaagactcaaaacttagcattgct accaaagagccacttacagtgttagatggaaaactggccctgcagacatcagcccccctctctgccactgataacaacgccctcactat cactgcctcacctcctcttactactgcaaatggtagtctggctgttaccatggaaaacccactttacaacaacaatggaaaacttgggctc aaaattggcggtcctttgcaagtggccaccgactcacatgcactaacactaggtactggtcagggggttgcagttcataacaatttgcta catacaaaagttacaggcgcaatagggtttgatacatctggcaacatggaacttaaaactggagatggcctctatgtggatagcgccgg tcctaaccaaaaactacatattaatctaaataccacaaaaggccttgcttttgacaacaccgcaataacaattaacgctggaaaagggttg gaatttgaaacagactcctcaaacggaaatcccataaaaacaaaaattggatcaggcatacaatataataccaatggagctatggttgc aaaacttggaacaggcctcagttttgacagctccggagccataacaatgggcagcataaacaatgacagacttactctttggacaacac cagacccatccccaaattgcagaattgcttcagataaagactgcaagctaactctggcgctaacaaaatgtggcagtcaaattttgggc actgtttcagctttggcagtatcaggtaatatggcctccatcaatggaactctaagcagtgtaaacttggttcttagatttgatgacaacgg agtgcttatgtcaaattcatcactggacaaacagtattggaactttagaaacggggactccactaacggtcaaccatacacttatgctgtt gggtttatgccaaacctaaaagcttacccaaaaactcaaagtaaaactgcaaaaagtaatattgttagccaggtgtatcttaatggtgaca agtctaaaccattgcattttactattacgctaaatggaacagatgaaaccaaccaagtaagcaaatactcaatatcattcagttggtcctgg aacagtggacaatacactaatgacaaatttgccaccaattcctataccttctcctacattgcccaggaataaagaatcgtgaacctgttgc atgttatgtttcaacgtgtttatttttcaattcgtattagtcatcgctattaccatggtgatgcggttttggcagtacatcaatgggcgtggatag cggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaat gtcgtaacaactccgccccattgacgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagagctggtttagtgaaccg tcagatccgctagagatctaaccggtggaaagcgctaccatgccttcatccgtgtcatggggaatcctgctgctggctggactgtgctgt ctggtgcctgtctcactggccgaggacccttttaatctggtgacaggctggcagactatcaatgggaagaaatactatttcgacattaac accggcgccgctctgatcagctacaagatcattaacgggaaacacttctacttcaacaatgacggagtgatgcagctgggcgtctttaa gggccccgatgggttcgagtacttcgcacctgccaatacacagaacaataacattgaaggacaggccatcgtgtatcagtccaaattc ctgactctgaacggcaagaaatactattttgaccaggattctaaggccgtcaccgggtggcgaatcattaataacgagaagtactacttc aaccccaataacgctattgcagccgtgggcctgcaggtcatcgacaataacaagtactatttcaaccctgatactgccatcatttccaaa ggatggcagaccgtgcagggctctcgctactatttcgacaccgatacagccatcgccttcaacgggtacaagaccatcgacggaaaa catttctattttgactcagattgcgtggtcaagatcggcgtgttcagcacctccaacggcttcgagtactttgccccagctaacacatacaa caacaacatcgagggccaggccatcgtgtaccagagcaagttcctgaccctgaatggcaagaaatactacttcgaccagaactctaa ggcagtcaccgggtggcagacaatcgatagtaagaagtactacttcaacactaacaccgccgaggctgcaactggctggcagaccat cgacgggaagaaatattatttcaatacaaacactgccgaagccgctacaggatggcagactattgacggcaagaaatattacttcaaca ccaacacagcaatcgcctctacagggtacactatcattaatggaaagcacttctacttcaacactgatgggatcatgcagattggagtgtt caaaggaccaaatggcttcgagtactttgctcccgcaaacacagacgccaacaatattgagggccaggctatcctgtatcagaatgaat tcctgacactgaacggcaagaaatattattttgggtctgatagtaaggctgtgactggctggaggatcattaacaataagaagtactattc aaccccaacaacgcaatcgcagccattcacctgtgcaccattaacaatgacaagtactacttcagctacgacggcatcctgcagaatg ggtatatcacaattgagcgcaacaatttctactttgacgccaaccaggaatccaagatggtgaccggcgtcttcaaagggcctaatgga tttgaatatttcgccccagctaacacacataataacaatatcgaggggcaggctatcgtgtatcagaataagttcctgaccctgaacggc aagaaatactactttgaccaggatagcaaagccgtgaccggatggcagacaatcgatggcaagaaatattatttcaatctgaacacagc cgaggccgcaactgggtggcagaccatcgatggaaagaaatactacttcaacctgaacactgctgaagccgctaccggatggcaga ctatcgacgggaagaaatactatttcaatactaacacctttattgcctctaccggatacacaagtatcaatggcaagcacttctacttcaac acggatggaatcatgcagattggcgtgttcaaaggccccaacggatttgaatactttgcacctgccaacactcataataacaatattgaa ggccaggctatcctgtaccaaaataagttcctgaccctgaacgggaagaaatattacttcggatcagacagcaaagccgtgaccggcc tgaggacaatcgatgggaagaaatattatttcaatacgaacactgctgtggcagtcactggatggcagaccattaatggcaagaaatatt acttcaacacgcagacaagcatcgcctccactgggtacaccatcattagcggaaagcacttctacttcaacaccgacggcattatgcag atcggagtgttcaaaggccctgatggatttgagtactttgcccccgctaatacagatgcaaataacattgaaggccaggccatccgata ccagaaccggttcctgtatctgcatgacaatatctactattttggccagaactccaaggcagccacaggctgggtgactatcgatggga atcggtactatttcgagcctaatacagctatgggggcaaacggatacaagactatcgataacaagaacttctacttccggaatggcctgc ctcagatcggggtgtttaagggcagcaacggattcgagtactttgcaccagccaacaccgacgccaataatattgaaggccaggcaat cagataccagaacaggttcctgcatctgctgggcaaaatctactacttcggccagaattccaaagcagtgactggctggcagacaatc aacggaaaggtctactacttcatgcctgacacagcaatggctgcagccggcggactgttcgagattgacggcgtgatctacttctttgg agtggatggcgtcaaagcacctggaatctacggaagaggacggagatcaagaggaaggcgcagcaacctgatcactggattcgtg accgtcggcgacgataagtactacttcaaccctattaacggaggcgctgcatccatcggcgagaccatcatcgacgataagaactact acttccagcagagtggggtgctgcagacaggagtcttctcaactgaggacggcttcaagtactttgctccagcaaataccctggatgaa aacctggagggagaagccattgactttacaggcaagctgatcatcgatgaaaacatctactacttcgacgataactaccgcggagctgt ggagtggaaagaactggacggcgagatgcactatttctctccagaaaccggcaaggccttcaaggggctgaatcagatcggagact acaagtactatttcaacagcgatggcgtgatgcagaaggggtttgtctccatcaatgacaacaaacactacttcgacgatagcggagtg atgaaggtcggctacaccgagattgatggcaaacatttctattttgctgagaatggggaaatgcaaatcggagtgttcaacacagaaga tggcttcaagtactttgcccaccataatgaggacctgggcaacgaggaaggggaggaaatttcctactctggcatcctgaacttcaaca acaaaatctactatttcgacgatagcttcaccgcagtggtgggatggaaggacctggaggatggaagcaaatactattttgacgaggat accgccgaagcttacattggcctgtccctgatcaatgacgggcagtactacttcaacgacgatggcattatgcaagtggggttcgtcac catcaacgacaaggtgttctactttagtgattcaggaatcattgagtctggcgtccagaatattgacgataactacttctatatcgacgata atgggatcgtgcagattggagtcttcgacaccagcgatgggtacaagtattttgcacccgccaacaccgtgaatgacaacatctacgg ccaggccgtcgagtattcaggcctggtgcgggtcggggaagacgtgtactatttcggcgagacttacaccattgaaacagggtggatc tatgacatggagcaagaaagtgataagtactatttcaatcctgagactaagaaagcctgcaaaggcatcaacctgattgacgatatcaa gtactacttcgatgagaagggaatcatgagaaccggcctgatcagcttcgaaaacaataactactacttcaacgagaacggggaaatg cagttcggatacatcaacatcgaggacaagatgttctacttcggggaagatggagtgatgcagatcggagtctttaacacacccgacg gcttcaaatactttgcccaccagaatactctggatgagaacttcgagggggaatctatccagtacaccggatggctggacctggatgag aagaggtactatttcaccgacgagtacatcgccgctacaggcagtgtgattatcgacggcgaggagtattacttcgatcccgacaccg ctcagctggtcatctcagagtaataaactagtaacggccgccagtgtgctggaattcgcccttataacttcgtatagcatacattatacga agttattgttgacaattaatcatcggcatagtatatcggcatagtataatacgacaaggtgaggaactaaaccatggccaagttgaccagt gccgttccggtgctcaccgcgcgcgacgtcgccggagcggtcgagttctggaccgaccggctcgggttctcccgggacttcgtgga ggacgacttcgccggtgtggtccgggacgacgtgaccctgttcatcagcgcggtccaggaccaggtggtgccggacaacaccctgg cctgggtgtgggtgcgcggcctggacgagctgtacgccgagtggtcggaggtcgtgtccacgaacttccgggacgcctccgggcc ggccatgaccgagatcggcgagcagccgtgggggcgggagttcgccctgcgcgacccggccggcaactgcgtgcacttcgtggc cgaggagcaggactgaataacttcgtatagcatacattatacgaagttataagggcgaattctgcagatatccatcctttaaaaaacctcc cacacctccccctgaacctgaaacataaaatgaatgcaattgttgttgttaacttgtttattgcagcttataatggttacaaataaagcaata gcatcacaaatttcacaaataaagcatttttttcactgcattctagttgtggtttgtccaaactcatcaatgtatcttaacaacgtgtttatttttc aattgcagaaagaattgcagaaaatttcaagtcatttttcattcagtagtatagccccaccaccacatagcttatactaatcaccgtacctta atcaaactcacagaaccctagtattcaacctgccacctccctcccaacacacagagtacacagtcctttctccccggctggccttaaaca gcatcatatcatgggtaacagacatattcttaggtgttatattccacacggtctcctgtcgagccaaacgctcatcagtgatgttaataaact ccccgggcagctcgcttaagttcatgtcgctgtccagctgctgagccacaggctgctgtccaacttgcggttgctcaacgggcggcga aggagaagtccacgcctacatgggggtagagtcataatcgtgcatcaggatagggcggtggtgctgcagcagcgcgcgaataaact gctgccgccgccgctccgtcctgcaggaatacaacatggcagtggtctcctcagcgatgattcgcaccgcccgcagcataaggcgc cttgtcctccgggcacagcagcgcaccctgatctcacttaagtcagcacagtaactgcagcacagtaccacaatattgtttaaaatccca cagtgcaaggcgctgtatccaaagctcatggcggggaccacagaacccacgtggccatcataccacaagcgcaggtagattaagtg gcgacccctcataaacacgctggacataaacattacctcttttggcatgttgtaattcaccacctcccggtaccatataaacctctgattaa acatggcgccatccaccaccatcctaaaccagctggccaaaacctgcccgccggctatgcactgcagggaaccgggactggaaca atgacagtggagagcccaggactcgtaaccatggatcatcatgctcgtcatgatatcaatgttggcacaacacaggcacacgtgcata cacttcctcaggattacaagctcctcccgcgtcagaaccatatcccagggaacaacccattcctgaatcagcgtaaatcccacactgca gggaagacctcgcacgtaactcacgttgtgcattgtcaaagtgttacattcgggcagcagcggatgatcctccagtatggtagcgcgtg tctctgtctcaaaaggaggtaggcgatccctactgtacggagtgcgccgagacaaccgagatcgtgttggtcgtagtgtcatgccaaat ggaacgccggacgtagtcatatttcctgaagcaaaaccaggtgcgggcgtgacaaacagatctgcgtctccggtctcgtcgcttagct cgctctgtgtagtagttgtagtatatccactctctcaaagcatccaggcgccccctggcttcgggttctatgtaaactccttcatgcgccgc tgccctgataacatccaccaccgcagaataagccacacccagccaacctacacattcgttctgcgagtcacacacgggaggagcgg gaagagctggaagaaccatgttttttttttttattccaaaagattatccaaaacctcaaaatgaagatctattaagtgaacgcgctcccctcc ggtggcgtggtcaaactctacagccaaagaacagataatggcatttgtaagatgttgcacaatggcttccaaaaggcaaactgccctca cgtccaagtggacgtaaaggctaaacccttcanggtgaatctcctctataaacattccagcaccttcaaccatgcccaaataattttcatct cgccaccttatcaatatgtctctaagcaaatcccgaatattaagtccggccattgtaaaaatctgctccagagcgccctccaccttcagcc tcaagcagcgaatcatgattgcaaaaattcaggttcctcacagacctgtataagattcaaaagcggaacattaacaaaaataccgcgat cccgtaggtcccttcgcagggccagctgaacataatcgtgcaggtctgcacggaccagcgcggccacttccccgccaggaaccatg acaaaagaacccacactgattatgacacgcatactcggagctatgctaaccagcgtagccccgatgtaagcttgttgcatgggcggcg atataaaatgcaaggtactgctcaaaaaatcaggcaaagcctcgcgcaaaaaagcaagcacatcgtagtcatgctcatgcagataaag gcaggtaagttccggaaccaccacagaaaaagacaccatttttctctcaaacatgtctgcgggttcctgcataaacacaaaataaaataa caaaaaaaaaaaaacatttaaacattagaagcctgtcttacaacaggaaaaacaacccttataagcataagacggactacggccatgc cggcgtgaccgtaaaaaaactggtcaccgtgattaaaaagcaccaccgacagttcctcggtcatgtccggagtcataatgtaagactc ggtaaacacatcaggttggttaacatcggtcagtgctaaaaagcgaccgaaatagcccgggggaatacatacccgcaggcgtagag acaacattacagcccccataggaggtataacaaaattaataggagagaaaaacacataaacccctgaaaaaccctcctgcccctagg caaaatagcaccctcccgctccagaacaacatacagcgcttccacagcggcagccataacagtcagccttaccagtaaaaaaacctat taaaaaacaccactcgacacggcaccagctcaatcagtcacagtgtaaaaagggccaagtacagagcgagtatatataggactaaaa aatgacgtaacggttaaagtccacaaaaaccacccagaaaaccgcacgcgaacctacgcccagaaacgaaagccaaaaaacccac aacttcctcaaatcttcacttccgttttcccacgatacgtcacttcccattttaaaaaaaaactacaattcccaatacatgcaagttactccgc cctaaaacctacgtcacccgccccgttcccacgccccgcgccacgtcacaaactccaccccctcattatcatattggcttcaatccaaa ataaggtatattattgatgatggcgat

[0494] SEQ ID NO:28 SC-Ad6-AIII-AE3-CMV-Spike-3X-LZL nucleic acid cgccatcatcaataatataccttattttggattgaagccaatatgataatgagggggtggagtttgtgacgtggcgcggggcgtgggaac ggggcgggtgacgtagtagtgtggcggaagtgtgatgttgtaagtgtggcggaacacatgtaagcgccggatgtggtaaaagtgacg tttttggtgtgcgccggtgtacacgggaagtgacaattttcgcgcggttttaggcggatgttgtagtaaatttgggcgtaaccaagtaatat ttggccattttcgcgggaaaactgaataagaggaagtgaaatctgaataattctgtgttactcatagcgcgtaatatttgtctagggccgc ggggactttgaccgtttacgtggagactcgcccaggtgtttttctcaggtgttttccgcgttccgggtcaaagttggcgttttattattatagt cagctgacgcgcagtgtatttatacccggtgagttcctcaagaggccactcttgagtgccagcgagtagagttttctcctccgagccgct ccgacaccgggactgaaaatgagacatattatctgccacggaggtgttattaccgaagaaatggccgccagtcttttggaccagctgat cgaagaggtactggctgataatcttccacctcctagccattttgaaccacctacccttcacgaactgtatgatttagacgtgacggccccc gaagatcccaacgaggaggcggtttcgcagatttttcccgagtctgtaatgttggcggtgcaggaagggattgacttattcacttttccgc cggcgcccggttctccggagccgcctcacctttcccggcagcccgagcagccggagcagagagccttgggtccggtttctatgcca aaccttgtgccggaggtgatcgatcttacctgccacgaggctggctttccacccagtgacgacgaggatgaagagggtgaggagtttg tgttagattatgtggagcaccccgggcacggttgcaggtcttgtcattatcaccggaggaatacgggggacccagatattatgtgttcgc tttgctatatgaggacctgtggcatgtttgtctacagtaagtgaaaaattatgggcagtgggtgatagagtggtgggtttggtgtggtaatt tttttttaatttttacagttttgtggtttaaagaattttgtattgtgattttttaaaaggtcctgtgtctgaacctgagcctgagcccgagccagaa ccggagcctgcaagacctacccggcgtcctaaattggtgcctgctatcctgagacgcccgacatcacctgtgtctagagaatgcaata gtagtacggatagctgtgactccggtccttctaacacacctcctgagatacacccggtggtcccgctgtgccccattaaaccagttgcc gtgagagttggtgggcgtcgccaggctgtggaatgtatcgaggacttgcttaacgagtctgggcaacctttggacttgagctgtaaacg ccccaggccataaggtgtaaacctgtgattgcgtgtgtggttaacgcctttgtttgctgaatgagttgatgtaagtttaataaagggtgaga taatgtttaacttgcatggcgtgttaaatggggcggggcttaaagggtatataatgcgccgtgggctaatcttggttacatctgacctcatg gaggcttgggagtgtttggaagatttttctgctgtgcgtaacttgctggaacagagctctaacagtacctcttggttttggaggtttctgtgg ggctcctcccaggcaaagttagtctgcagaattaaggaggattacaagtgggaatttgaagagcttttgaaatcctgtggtgagctgtttg attctttgaatctgggtcaccaggcgcttttccaagagaaggtcatcaagactttggatttttccacaccggggcgcgctgcggctgctgt tgcttttttgagttttataaaggataaatggagcgaagaaacccatctgagcggggggtacctgctggattttctggccatgcatctgtgg agagcggtggtgagacacaagaatcgcctgctactgttgtcttccgtccgcccggcaataataccgacggaggagcaacagcagga ggaagccaggcggcggcggcggcaggagcagagcccatggaacccgagagccggcctggaccctcgggaatgaatgttgtaca ggtggctgaactgtttccagaactgagacgcattttaaccattaacgaggatgggcaggggctaaagggggtaaagagggagcggg gggcttctgaggctacagaggaggctaggaatctaacttttagcttaatgaccagacaccgtcctgagtgtgttacttttcagcagattaa ggataattgcgctaatgagcttgatctgctggcgcagaagtattccataaagcagctgaccacttactggctgcagccaggggatgattt tgaggaggctattagggtatatgcaaaggtggcacttaggccagattgcaagtacaagattagcaaacttgtaaatatcaggaattgttg ctacatttctgggaacggggccgaggtggagatagatacggaggatagggtggcctttagatgtagcatgataaatatgtggccggg ggtgcttggcatggacggggtggttattatgaatgtgaggtttactggtcccaattttagcggtacggttttcctggccaataccaatcttat cctacacggtgtaagcttctatgggtttaacaatacctgtgtggaagcctggaccgatgtaagggttcggggctgtgccttttactgctgc tggaagggggtggtgtgtcgccccaaaagcagggcttcaattaagaaatgcctgtttgaaaggtgtaccttgggtatcctgtctgaggg taactccagggtgcgccacaatgtggcctccgactgtggttgctttatgctagtgaaaagcgtggctgtgattaagcataacatggtgtgt ggcaactgcgaggacagggcctctcagatgctgacctgctcggacggcaactgtcacttgctgaagaccattcacgtagccagccac tctcgcaaggcctggccagtgtttgagcacaacatactgacccgctgttccttgcatttgggtaacaggaggggggtgttcctaccttac caatgcaatttgagtcacactaagatattgcttgagcccgagagcatgtccaaggtgaacctgaacggggtgtttgacatgaccatgaa gatctggaaggtgctgaggtacgatgagacccgcaccaggtgcagaccctgcgagtgtggcggtaaacatattaggaaccagcctgt gatgctggatgtgaccgaggagctgaggcccgatcacttggtgctggcctgcacccgcgctgagtttggctctagcgatgaagataca gattgaggtactgaaatgtgtgggcgtggcttaagggtgggaaagaatatataaggtgggggtctcatgtagttttgtatctgttttgcag cagccgccgccatgagcgccaactcgtttgatggaagcattgtgagctcatatttgacaacgcgcatgcccccatgggccggggtgc gtcagaatgtgatgggctccagcattgatggtcgccccgtcctgcccgcaaactctactaccttgacctacgagaccgtgtctggaacg ccgttggagactgcagcctccgccgccgcttcagccgctgcagccaccgcccgcgggattgtgactgactttgctttcctgagcccgc ttgcaagcagtgcagcttcccgttcatccgcccgcgatgacaagttgacggctcttttggcacaattggattctttgacccgggaacttaa tgtcgtttctcagcagctgttggatctgcgccagcaggtttctgccctgaaggcttcctcccctcccaatgcggtttaaaacataaataaaa accagactctgtttggatttggatcaagcaagtgtcttgctgtctttatttaggggttttgcgcgcgcggtaggcccgggaccagcggtct cggtcgttgagggtcctgtgtattttttccaggacgtggtaaaggtgactctggatgttcagatacatgggcataagcccgtctctggggt ggaggtagcaccactgcagagcttcatgctgcggggtggtgttgtagatgatccagtcgtagcaggagcgctgggcgtggtgcctaa aaatgtctttcagtagcaagctgattgccaggggcaggcccttggtgtaagtgtttacaaagcggttaagctgggatgggtgcatacgt ggggatatgagatgcatcttggactgtatttttaggttggctatgttcccagccatatccctccggggattcatgttgtgcagaaccaccag cacagtgtatccggtgcacttgggaaatttgtcatgtagcttagaaggaaatgcgtggaagaacttggagacgcccttgtgacctccaa gattttccatgcattcgtccataatgatggcaatgggcccacgggcggcggcctgggcgaagatatttctgggatcactaacgtcatagt tgtgttccaggatgagatcgtcataggccatttttacaaagcgcgggcggagggtgccagactgcggtataatggttccatccggccca ggggcgtagttaccctcacagatttgcatttcccacgctttgagttcagatggggggatcatgtctacctgcggggcgatgaagaaaac cgtttccggggtaggggagatcagctgggaagaaagcaggttcctaagcagctgcgacttaccgcagccggtgggcccgtaaatca cacctattaccggctgcaactggtagttaagagagctgcagctgccgtcatccctgagcaggggggccacttcgttaagcatgtccctg acttgcatgttttccctgaccaaatccgccagaaggcgctcgccgcccagcgatagcagttcttgcaaggaagcaaagtttttcaacgg tttgaggccgtccgccgtaggcatgcttttgagcgtttgaccaagcagttccaggcggtcccacagctcggtcacgtgctctacggcat ctcgatccagcatatctcctcgtttcgcgggttggggcggctttcgctgtacggcagtagtcggtgctcgtccagacgggccagggtca tgtctttccacgggcgcagggtcctcgtcagcgtagtctgggtcacggtgaaggggtgcgctccgggttgcgcgctggccagggtgc gcttgaggctggtcctgctggtgctgaagcgctgccggtcttcgccctgcgcgtcggccaggtagcatttgaccatggtgtcatagtcc agcccctccgcggcgtggcccttggcgcgcagcttgcccttggaggaggcgccgcacgaggggcagtgcagacttttaagggcgt agagcttgggcgcgagaaataccgattccggggagtaggcatccgcgccgcaggccccgcagacggtctcgcattccacgagcca ggtgagctctggccgttcggggtcaaaaaccaggtttcccccatgctttttgatgcgtttcttacctctggtttccatgagccggtgtccac gctcggtgacgaaaaggctgtccgtgtccccgtatacagacttgagaggcctgtcctcgagcggtgttccgcggtcctcctcgtataga aactcggaccactctgagacgaaggctcgcgtccaggccagcacgaaggaggctaagtgggaggggtagcggtcgttgtccacta gggggtccactcgctccagggtgtgaagacacatgtcgccctcttcggcatcaaggaaggtgattggtttataggtgtaggccacgtg accgggtgttcctgaaggggggctataaaagggggtgggggcgcgttcgtcctcactctcttccgcatcgctgtctgcgagggccag ctgttggggtgagtactccctctcaaaagcgggcatgacttctgcgctaagattgtcagtttccaaaaacgaggaggatttgatattcacc tggcccgcggtgatgcctttgagggtggccgcgtccatctggtcagaaaagacaatctttttgttgtcaagcttggtggcaaacgaccc gtagagggcgttggacagcaacttggcgatggagcgcagggtttggtttttgtcgcgatcggcgcgctccttggccgcgatgtttagct gcacgtattcgcgcgcaacgcaccgccattcgggaaagacggtggtgcgctcgtcgggcactaggtgcacgcgccaaccgcggtt gtgcagggtgacaaggtcaacgctggtggctacctctccgcgtaggcgctcgttggtccagcagaggcggccgcccttgcgcgagc agaatggcggtagtgggtctagctgcgtctcgtccggggggtctgcgtccacggtaaagaccccgggcagcaggcgcgcgtcgaa gtagtctatcttgcatccttgcaagtctagcgcctgctgccatgcgcgggcggcaagcgcgcgctcgtatgggttgagtgggggaccc catggcatggggtgggtgagcgcggaggcgtacatgccgcaaatgtcgtaaacgtagaggggctctctgagtattccaagatatgta gggtagcatcttccaccgcggatgctggcgcgcacgtaatcgtatagttcgtgcgagggagcgaggaggtcgggaccgaggttgct acgggcgggctgctctgctcggaagactatctgcctgaagatggcatgtgagttggatgatatggttggacgctggaagacgttgaag ctggcgtctgtgagacctaccgcgtcacgcacgaaggaggcgtaggagtcgcgcagcttgttgaccagctcggcggtgacctgcac gtctagggcgcagtagtccagggtttccttgatgatgtcatacttatcctgtcccttttttttccacagctcgcggttgaggacaaactcttc gcggtctttccagtactcttggatcggaaacccgtcggcctccgaacggtaagagcctancatgtagaactggttgacggcctggtag gcgcagcatcccttttctacgggtagcgcgtatgcctgcgcggccttccggagcgaggtgtgggtgagcgcaaaggtgtccctaacc atgactttgaggtactggtatttgaagtcagtgtcgtcgcatccgccctgctcccagagcaaaaagtccgtgcgctttttggaacgcggg tttggcagggcgaaggtgacatcgttgaagagtatctttcccgcgcgaggcataaagttgcgtgtgatgcggaagggtcccggcacct cggaacggttgttaattacctgggcggcgagcacgatctcgtcaaagccgttgatgttgtggcccacaatgtaaagttccaagaagcg cgggatgcccttgatggaaggcaattttttaagttcctcgtaggtgagctcttcaggggagctgagcccgtgctctgaaagggcccagt ctgcaagatgagggttggaagcgacgaatgagctccacaggtcacgggccattagcatttgcaggtggtcgcgaaaggtcctaaact ggcgacctatggccattttttctggggtgatgcagtagaaggtaagcgggtcttgttcccagcggtcccatccaaggtccgcggctagg tctcgcgcggcggtcactagaggctcatctccgccgaacttcatgaccagcatgaagggcacgagctgcttcccaaaggcccccatc caagtataggtctctacatcgtaggtgacaaagagacgctcggtgcgaggatgcgagccgatcgggaagaactggatctcccgcca ccagttggaggagtggctgttgatgtggtgaaagtagaagtccctgcgacgggccgaacactcgtgctggcttttgtaaaaacgtgcg cagtactggcagcggtgcacgggctgtacatcctgcacgaggttgacctgacgaccgcgcacaaggaagcagagtgggaatttgag cccctcgcctggcgggtttggctggtggtcttctacttcggctgcttgtccttgaccgtctggctgctcgaggggagttacggtggatcg gaccaccacgccgcgcgagcccaaagtccagatgtccgcgcgcggcggtcggagcttgatgacaacatcgcgcagatgggagct gtccatggtctggagctcccgcggcgtcaggtcaggcgggagctcctgcaggtttacctcgcatagccgggtcagggcgcgggcta ggtccaggtgatacctgatttccaggggctggttggtggcggcgtcgatggcttgcaagaggccgcatccccgcggcgcgactacg gtaccgcgcggcgggcggtgggccgcgggggtgtccttggatgatgcatctaaaagcggtgacgcgggcgggcccccggaggta gggggggctcgggacccgccgggagagggggcaggggcacgtcggcgccgcgcgcgggcaggagctggtgctgcgcgcgg aggttgctggcgaacgcgacgacgcggcggttgatctcctgaatctggcgcctctgcgtgaagacgacgggcccggtgagcttgaa cctgaaagagagttcgacagaatcaatttcggtgtcgttgacggcggcctggcgcaaaatctcctgcacgtctcctgagttgtcttgata ggcgatctcggccatgaactgctcgatctcttcctcctggagatctccgcgtccggctcgctccacggtggcggcgaggtcgttggag atgcgggccatgagctgcgagaaggcgttgaggcctccctcgttccagacgcggctgtagaccacgcccccttcggcatcgcgggc gcgcatgaccacctgcgcgagattgagctccacgtgccgggcgaagacggcgtagtttcgcaggcgctgaaagaggtagttgagg gtggtggcggtgtgttctgccacgaagaagtacataacccagcgccgcaacgtggattcgttgatatcccccaaggcctcaaggcgct ccatggcctcgtagaagtccacggcgaagttgaaaaactgggagttgcgcgccgacacggttaactcctcctccagaagacggatga gctcggcgacagtgtcgcgcacctcgcgctcaaaggctacaggggcctcttcttcttcttcaatctcctcttccataagggcctccccttc ttcttcttctggcggcggtgggggaggggggacacggcggcgacgacggcgcaccgggaggcggtcgacaaagcgctcgatcat ctccccgcggcgacggcgcatggtctcggtgacggcgcggccgttctcgcgggggcgcagttggaagacgccgcccgtcatgtcc cggttatgggttggcggggggctgccgtgcggcagggatacggcgctaacgatgcatctcaacaattgttgtgtaggtactccgccac cgagggacctgagcgagtccgcatcgaccggatcggaaaacctctcgagaaaggcgtctaaccagtcacagtcgcaaggtaggct gagcaccgtggcgggcggcagcgggcggcggtcggggttgtttctggcggaggtgctgctgatgatgtaattaaagtaggcggtctt gagacggcggatggtcgacagaagcaccatgtccttgggtccggcctgctgaatgcgcaggcggtcggccatgccccaggcttcgt tttgacatcggcgcaggtctttgtagtagtcttgcatgagcctttctaccggcacttcttcttctccttcctcttgtcctgcatctcttgcatctat cgctgcggcggcggcggagtttggccgtaggtggcgccctcttcctcccatgcgtgtgaccccgaagcccctcatcggctgaagca gggccaggtcggcgacaacgcgctcggctaatatggcctgctgcacctgcgtgagggtagactggaagtcgtccatgtccacaaag cggtggtatgcgcccgtgttgatggtgtaagtgcagttggccataacggaccagttaacggtctggtgacccggctgcgagagctcgg tgtacctgagacgcgagtaagcccttgagtcaaagacgtagtcgttgcaagtccgcaccaggtactggtatcccaccaaaaagtgcgg cggcggctggcggtagaggggccagcgtagggtggccggggctccgggggcgaggtcttccaacataaggcgatgatatccgta gatgtacctggacatccaggtgatgccggcggcggtggtggaggcgcgcggaaagtcacggacgcggttccagatgttgcgcagc ggcaaaaagtgctccatggtcgggacgctctggccggtcaggcgcgcgcagtcgttgacgctctagaccgtgcaaaaggagagcct gtaagcgggcactcttccgtggtctggtggataaattcgcaagggtatcatggcggacgaccggggttcgaaccccggatccggccg tccgccgtgatccatgcggttaccgcccgcgtgtcgaacccaggtgtgcgacgtcagacaacgggggagcgctccttttggcttcctt ccaggcgcggcggatgctgcgctagcttttttggccactggccgcgcgcggcgtaagcggttaggctggaaagcgaaagcattaagt ggctcgctccctgtagccggagggttattttccaagggttgagtcgcgggacccccggttcgagtctcgggccggccggactgcggc gaacgggggtttgcctccccgtcatgcaagaccccgcttgcaaattcctccggaaacagggacgagccccttttttgcttttcccagatg catccggtgctgcggcagatgcgcccccctcctcagcagcggcaagagcaagagcagcggcagacatgcagggcaccctcccct cctcctaccgcgtcaggaggggcgacatccgcggttgacgcggcagcagatggtgattacgaacccccgcggcgccgggcccgg cactacctggacttggaggagggcgagggcctggcgcggctaggagcgccctctcctgagcggtacccaagggtgcagctgaagc gtgatacgcgtgaggcgtacgtgccgcggcagaacctgtttcgcgaccgcgagggagaggagcccgaggagatgcgggatcgaa agttccacgcagggcgcgagctgcggcatggcctgaatcgcgagcggttgctgcgcgaggaggactttgagcccgacgcgcgaac cgggattagtcccgcgcgcgcacacgtggcggccgccgacctggtaaccgcatacgagcagacggtgaaccaggagattaactttc aaaaaagctttaacaaccacgtgcgtacgcttgtggcgcgcgaggaggtggctataggactgatgcatctgtgggactttgtaagcgc gctggagcaaaacccaaatagcaagccgctcatggcgcagctgttccttatagtgcagcacagcagggacaacgaggcattcaggg atgcgctgctaaacatagtagagcccgagggccgctggctgctcgatttgataaacatcctgcagagcatagtggtgcaggagcgca gcttgagcctggctgacaaggtggccgccatcaactattccatgcttagcctgggcaagttttacgcccgcaagatataccatacccctt acgttcccatagacaaggaggtaaagatcgaggggttctacatgcgcatggcgctgaaggtgcttaccttgagcgacgacctgggcg tttatcgcaacgagcgcatccacaaggccgtgagcgtgagccggcggcgcgagctcagcgaccgcgagctgatgcacagcctgca aagggccctggctggcacgggcagcggcgatagagaggccgagtcctactttgacgcgggcgctgacctgcgctgggccccaag ccgacgcgccctggaggcagctggggccggacctgggctggcggtggcacccgcgcgcgctggcaacgtcggcggcgtggag gaatatgacgaggacgatgagtacgagccagaggacggcgagtactaagcggtgatgtttctgatcagtcgcggccgcgatatcgct agcgaagttcctattctctagaaagtataggaacttcggatcctctagagtcgaaaaaaaaaaagcatgatgcaaaataaaaaactcac caaggccatggcaccgagcgttggttttcttgtattccccttagtatgcggcgcgcggcgatgtatgaggaaggtcctcctccctcctac gagagtgtggtgagcgcggcgccagtggcggcggcgctgggttctcccttcgatgctcccctggacccgccgtttgtgcctccgcgg tacctgcggcctaccggggggagaaacagcatccgttactctgagttggcacccctattcgacaccacccgtgtgtacctggtggaca acaagtcaacggatgtggcatccctgaactaccagaacgaccacagcaactttctgaccacggtcattcaaaacaatgactacagccc gggggaggcaagcacacagaccatcaatcttgacgaccggtcgcactggggcggcgacctgaaaaccatcctgcataccaacatg ccaaatgtgaacgagttcatgtttaccaataagtttaaggcgcgggtgatggtgtcgcgcttgcctactaaggacaatcaggtggagct gaaatacgagtgggtggagttcacgctgcccgagggcaactactccgagaccatgaccatagaccttatgaacaacgcgatcgtgga gcactacttgaaagtgggcagacagaacggggttctggaaagcgacatcggggtaaagtttgacacccgcaacttcagactggggtt tgaccccgtcactggtcttgtcatgcctggggtatatacaaacgaagccttccatccagacatcattttgctgccaggatgcggggtgga cttcacccacagccgcctgagcaacttgttgggcatccgcaagcggcaacccttccaggagggctttaggatcacctacgatgatctg gagggtggtaacattcccgcactgttggatgtggacgcctaccaggcgagcttgaaagatgacaccgaacagggcgggggtggcg caggcggcagcaacagcagtggcagcggcgcggaagagaactccaacgcggcagccgcggcaatgcagccggtggaggacat gaacgatcatgccattcgcggcgacacctttgccacacgggctgaggagaagcgcgctgaggccgaagcagcggccgaagctgc cgcccccgctgcgcaacccgaggtcgagaagcctcagaagaaaccggtgatcaaacccctgacagaggacagcaagaaacgcag ttacaacctaataagcaatgacagcaccttcacccagtaccgcagctggtaccttgcatacaactacggcgaccctcagaccggaatc cgctcatggaccctgctttgcactcctgacgtaacctgcggctcggagcaggtctactggtcgttgccagacatgatgcaagaccccgt gaccttccgctccacgcgccagatcagcaactttccggtggtgggcgccgagctgttgcccgtgcactccaagagcttctacaacgac caggccgtctactcccaactcatccgccagtttacctctctgacccacgtgttcaatcgctttcccgagaaccagattttggcgcgcccg ccagcccccaccatcaccaccgtcagtgaaaacgttcctgctctcacagatcacgggacgctaccgctgcgcaacagcatcggagg agtccagcgagtgaccattactgacgccagacgccgcacctgcccctacgtttacaaggccctgggcatagtctcgccgcgcgtcct atcgagccgcactttttgagcaagcatgtccatccttatatcgcccagcaataacacaggctggggcctgcgcttcccaagcaagatgtt tggcggggccaagaagcgctccgaccaacacccagtgcgcgtgcgcgggcactaccgcgcgccctggggcgcgcacaaacgcg gccgcactgggcgcaccaccgtcgatgacgccatcgacgcggtggtggaggaggcgcgcaactacacgcccacgccgccgcca gtgtccaccgtggacgcggccattcagaccgtggtgcgcggagcccggcgctacgctaaaatgaagagacggcggaggcgcgta gcacgtcgccaccgccgccgacccggcactgccgcccaacgcgcggcggcggccctgcttaaccgcgcacgtcgcaccggccg acgggcggccatgcgagccgctcgaaggctggccgcgggtattgtcactgtgccccccaggtccaggcgacgagcggccgccgc agcagccgcggccattagtgttatgactcagggtcgcaggggcaacgtgtactgggtgcgcgactcggttagcggcctgcgcgtgc ccgtgcgcacccgccccccgcgcaactagattgcaataaaaaactacttagactcgtactgttgtatgtatccagcggcggcggcgcg catcgaagctatgtccaagcgcaaaatcaaagaagagatgctccaggtcatcgcgccggagatctatggccccccgaagaaggaag agcaggattacaagccccgaaagctaaagcgggtcaaaaagaaaaagaaagatgatgatgatgatgaacttgacgacgaggtggaa ctgttgcacgcgaccgcgcccaggcgacgggtacagtggaaaggtcgacgcgtaagacgtgttttgcgacccgg caeca ccgtagt ctttacgcccggtgagcgctccacccgcacctacaagcgcgtgtatgatgaggtgtacggcgacgaggacctgcttgagcaggcca acgagcgcctcggggagtttgcctacggaaagcggcataaggacatgctggcgttgccgctggacgagggcaacccaacacctag cctaaagcccgtgacactgcagcaggtgctgcccgcgcttgcaccgtccgaagaaaagcgcggcctaaagcgcgagtctggtgact tggcacccaccgtgcagctgatggtacccaagcgtcagcgactggaagatgtcttggaaaaaatgaccgtggagcctgggctggag cccgaggtccgcgtgcggccaatcaagcaggtggcaccgggactgggcgtgcagaccgtggacgttcagatacccaccaccagta gcactagtattgccactgccacagagggcatggagacacaaacgtccccggttgcctcggcggtggcagatgccgcggtgcaggc ggccgctgcggccgcgtccaagacctctacggaggtgcaaacggacccgtggatgtttcgtgtttcagccccccggcgtccgcgcc gttcaaggaagtacggcgccgccagcgcgctactgcccgaatatgccctacatccttccatcgcgcctacccccggctatcgtggcta cacctaccgccccagaagacgagcaactacccgacgccgaaccaccactggaacccgccgccgccgtcgccgtcgccagcccgt gctggccccgatttccgtgcgcagggtggctcgcgaaggaggcaggaccctggtgctgccaacagcgcgctaccaccccagcatc gtttaaaagccggtctttgtggttcttgcagatatggccctcacctgccgcctccgtttcccggtgccgggattccgaggaagaatgcac cgtaggaggggcatggccggccacggcctgacgggcggcatgcgtcgtgcgcaccaccggcggcggcgcgcgtcgcaccgtcg catgcgcgccggtatcctgcccctccttattccactgatcgccgcggcgattggcgccgtgcccggaattgcatccgtggccttgcag gcgcagagacactgattaaaaacaagttacatgtggaaaaatcaaaataaaagtctggactctcacgctcgcttggtcctgtaactatttt gtagaatggaagacatcaactttgcgtcactggccccgcgacacggctcgcgcccgttcatgggaaactggcaagatatcgg caeca gcaatatgagcggtggcgccttcagctggggctcgctgtggagcggcattaaaaatttcggttccgccgttaagaactatggcagcaa agcctggaacagcagcacaggccagatgctgagggacaagttgaaagagcaaaatttccaacaaaaggtggtagatggcctggcct ctggcattagcggggtggtggacctggccaaccaggcagtgcaaaataagattaacagtaagcttgatccccgccctcccgtagagg agcctccaccggccgtggagacagtgtctccagaggggcgtggcgaaaagcgtccgcgacccgacagggaagaaactctggtga cgcaaatagacgagcctccctcgtacgaggaggcactaaagcaaggcctgcccaccacccgtcccatcgcgcccatggctaccgg agtgctgggccagcacacacccgtaacgctggacctgcctccccccgccgacacccagcagaaacctgtgctgccaggcccgtcc gccgttgttgtaacccgtcctagccgcgggtccctgcgccgcgccgccagcggtccgcgatcgttgcggcccgtagccagtggcaa ctggcaaagcacactgaacagcatcgtgggtttgggggtgcaatccctgaagcgccgacgatgcttctgatagctaacgtgtcgtatgt gtgtcatgtatgcgtccatgtcgccgccagaggagctgctgagccgccgcgcgcccgctttccaagatggctaccccttcgatgatgc cgcagtggtcttacatgcacatctcgggccaggacgcctcggagtacctgagccccgggctggtgcagttcgcccgcgccaccgag acgtacttcagcctgaataacaagtttagaaaccccacggtggcgcctacgcacgacgtgaccacagaccggtctcagcgtttgacgc tgcggttcatccccgtggaccgcgaggatactgcgtactcgtacaaggcgcggttcaccctagctgtgggtgataaccgtgtgctaga catggcttccacgtactttgacatccgcggcgtgctggacaggggccctacttttaagccctactctggcactgcctacaacgcactgg cccccaagggtgcccccaactcgtgcgagtgggaacaaaatgaaactgcacaagtggatgctcaagaacttgacgaagaggagaat gaagccaatgaagctcaggcgcgagaacaggaacaagctaagaaaacccatgtatatgcccaggctccactgtccggaataaaaat aactaaagaaggtctacaaataggaactgccgacgccacagtagcaggtgccggcaaagaaattttcgcagacaaaacttttcaacct gaaccacaagtaggagaatctcaatggaacgaagcggatgccacagcagctggtggaagggttcttaaaaagacaactcccatgaa accctgctatggctcatacgctagacccaccaattccaacggcggacagggcgttatggttgaacaaaatggtaaattggaaagtcaa gtcgaaatgcaatttttttccacatccacaaatgccacaaatgaagttaacaatatacaaccaacagttgtattgtacagcgaagatgtaaa catggaaactccagatactcatctttcttataaacctaaaatgggggataaaaatgccaaagtcatgcttggacaacaagcaatgccaaa cagaccaaattacattgcttttagagacaattttattggtctcatgtattacaacagcacaggtaacatgggtgtccttgctggtcaggcatc gcagttgaacgctgttgtagatttgcaagacagaaacacagagctgtcctaccagcttttgcttgattcaattggcgacagaacaagata cttttcaatgtggaatcaagctgttgacagctatgatccagatgtcagaattattgagaaccatggaactgaggatgagttgccaaattatt gctttcctcttggtggaattgggattactgacacttttcaagctgttaaaacaactgctgctaacggggaccaaggcaatactacctggca aaaagattcaacatttgcagaacgcaatgaaataggggtgggaaataactttgccatggaaattaacctgaatgccaacctatggagaa atttcctttactccaatattgcgctgtacctgccagacaagctaaaatacaaccccaccaatgtggaaatatctgacaaccccaacaccta cgactacatgaacaagcgagtggtggctcctgggcttgtagactgctacattaaccttggggcgcgctggtctctggactacatggaca acgttaatccctttaaccacccccgccatgcgggcctgcgttaccgctccatgttgttgggaaacggccgctacgtgccctttcacattc aggtgccccaaaagttttttgccattaaaaacctcctcctcctgccaggctcatacacatatgaatggaacttcaggaaggatgttaacat ggttctgcagagctctctgggaaacgaccttagagttgacggggctagcattaagtttgacagcatttgtctttacgccaccttcttcccca tggcccacaacacggcctccacgctggaagccatgctcagaaatgacaccaacgaccagtcctttaatgactacctttccgccgccaa catgctatatcccatacccgccaacgccaccaacgtgcccatctccatcccatcgcgcaactgggcagcatttcgcggttgggccttca cacgcttgaagacaaaggaaaccccttccctgggatcaggctacgacccttactacacctactctggctccataccataccttgacgga accttctatcttaatcacacctttaagaaggtggccattacttttgactcttctgttagctggccgggcaacgaccgcctgcttactcccaat gagtttgagattaagcgctcagttgacggggagggctataacgtagctcagtgcaacatgacaaaggactggttcctagtgcagatgtt ggccaactacaatattggctaccagggcttctacattccagaaagctacaaagaccgcatgtactcgttcttcagaaacttccagcccat gagccggcaagtggtggacgatactaaatacaaagattatcagcaggttggaattatccaccagcataacaactcaggcttcgtaggct acctcgctcccaccatgcgcgagggacaagcttaccccgctaatgttccctacccactaataggcaaaaccgcggttgatagtattacc cagaaaaagtttctttgcgaccgcaccctgtggcgcatccccttctccagtaactttatgtccatgggtgcgctcacagacctgggccaa aaccttctctacgcaaactccgcccacgcgctagacatgacctttgaggtggatcccatggacgagcccacccttctttatgttttgtttga agtctttgacgtggtccgtgtgcaccagccgcaccgcggcgtcatcgagaccgtgtacctgcgcacgcccttctcggccggcaacgc cacaacataaagaagcaagcaacatcaacaacagctgccgccatgggctccagtgagcaggaactgaaagccattgtcaaagatctt ggttgtgggccatattttttgggcacctatgacaagcgcttcccaggctttgtttccccacacaagctcgcctgcgccatagttaacacgg ccggtcgcgagactgggggcgtacactggatggcctttgcctggaacccgcgctcaaaaacatgctacctctttgagccctttggctttt ctgaccaacgtctcaagcaggtttaccagtttgagtacgagtcactcctgcgccgtagcgccattgcctcttcccccgaccgctgtataa cgctggaaaagtccacccaaagcgtgcaggggcccaactcggccgcctgtggcctattctgctgcatgtttctccacgcctttgccaac tggccccaaactcccatggatcacaaccccaccatgaaccttattaccggggtacccaactccatgcttaacagtccccaggtacagc ccaccctgcgccgcaaccaggaacagctctacagcttcctggagcgccactcgccctacttccgcagccacagtgcgcaaattagga gcgccacttctttttgtcacttgaaaaacatgtaaaaataatgtactaggagacactttcaataaaggcaaatgtttttatttgtacactctcg ggtgattatttacccccacccttgccgtctgcgccgtttaaaaatcaaaggggttctgccgcgcatcgctatgcgccactggcagggac acgttgcgatactggtgtttagtgctccacttaaactcaggcacaaccatccgcggcagctcggtgaagttttcactccacaggctgcgc accatcaccaacgcgtttagcaggtcgggcgccgatatcttgaagtcgcagttggggcctccgccctgcgcgcgcgagttgcgatac acagggttacagcactggaacactatcagcgccgggtggtgcacgctggccagcacgctcttgtcggagatcanatccgcgtccag gtcctccgcgttgctcagggcgaacggagtcaactttggtagctgccttcccaaaaagggtgcatgcccaggctttgagttgcactcgc accgtagtggcatcagaaggtgaccgtgcccagtctgggcgttaggatacagcgcctgcatgaaagccttgatctgcttaaaagccac ctgagcctttgcgccttcagagaagaacatgccgcaagacttgccggaaaactgattggccggacaggccgcgtcatgcacgcagc accttgcgtcggtgttggagatctgcaccacatttcggccccaccggttcttcacgatcttggccttgctagactgctccttcagcgcgcg ctgcccgttttcgctcgtcacatccatttcaatcacgtgctccttatttatcataatgctcccgtgtagacacttaagctcgccttcgatctca gcgcagcggtgcagccacaacgcgcagcccgtgggctcgtggtgcttgtaggttacctctgcaaacgactgcaggtacgcctgcag gaatcgccccatcatcgtcacaaaggtcttgttgctggtgaaggtcagctgcaacccgcggtgctcctcgtttagccaggtcttgcatac ggccgccagagcttccacttggtcaggcagtagcttgaagtttgcctttagatcgttatccacgtggtacttgtccatcaacgcgcgcgc agcctccatgcccttctcccacgcagacacgatcggcaggctcagcgggtttatcaccgtgctttcactttccgcttcactggactcttcc ttttcctcttgcatccgcataccccgcgccactgggtcgtcttcattcagccgccgcaccgtgcgcttacctcccttgccgtgcttgattag caccggtgggttgctgaaacccaccatttgtagcgccacatcttctctttcttcctcgctgtccacgatcacctctggggatggcgggcg ctcgggcttgggagaggggcgcttctttttctttttggacgcaatggccaaatccgccgtcgaggtcgatggccgcgggctgggtgtgc gcggcaccagcgcatcttgtgacgagtcttcttcgtcctcggactcgagacgccgcctcagccgcttttttgggggcgcgcggggag gcggcggcgacggcgacggggacgagacgtcctccatggttggtggacgtcgcgccgcaccgcgtccgcgctcgggggtggttt cgcgctgctcctcttcccgactggccatttccttctcctataggcagaaaaagatcatggagtcagtcgagaaggaggacagcctaacc gccccctttgagttcgccaccaccgcctccaccgatgccgccaacgcgcctaccaccttccccgtcgaggcacccccgcttgaggag gaggaagtgattatcgagcaggacccaggttttgtaagcgaagacgacgaagatcgctcagtaccaacagaggataaaaagcaaga ccaggacgacgcagaggcaaacgaggaacaagtcgggcggggggaccaaaggcatggcgactacctagatgtgggagacgac gtgctgttgaagcatctgcagcgccagtgcgccattatctgcgacgcgttgcaagagcgcagcgatgtgcccctcgccatagcggat gtcagccttgcctacgaacgccacctgttctcaccgcgcgtaccccccaaacgccaagaaaacggcacatgcgagcccaacccgcg cctcaacttctaccccgtatttgccgtgccagaggtgcttgccacctatcacatctttttccaaaactgcaagatacccctatcctgccgtg ccaaccgcagccgagcggacaagcagctggccttgcggcagggcgctgtcatacctgatatcgcctcgctcgacgaagtgccaaaa atctttgagggtcttggacgcgacgagaagcgcgcggcaaacgctctgcaacaagaaaacagcgaaaatgaaagtcactgtggagt gctggtggaacttgagggtgacaacgcgcgcctagccgtgctgaaacgcagcatcgaggtcacccactttgcctacccggcacttaa cctaccccccaaggttatgagcacagtcatgagcgagctgatcgtgcgccgtgcacgacccctggagagggatgcaaacttgcaag aacaaaccgaggagggcctacccgcagttggcgatgagcagctggcgcgctggcttgagacgcgcgagcctgccgacttggagg agcgacgcaagctaatgatggccgcagtgcttgttaccgtggagcttgagtgcatgcagcggttctttgctgacccggagatgcagcg caagctagaggaaacgttgcactacacctttcgccagggctacgtgcgccaggcctgcaaaatttccaacgtggagctctgcaacctg gtctcctaccttggaattttgcacgaaaaccgccttgggcaaaacgtgcttcattccacgctcaagggcgaggcgcgccgcgactacg tccgcgactgcgtttacttatttctgtgctacacctggcaaacggccatgggcgtgtggcagcagtgcctggaggagcgcaacctgaa ggagctgcagaagctgctaaagcaaaacttgaaggacctatggacggccttcaacgagcgctccgtggccgcgcacctggcggac attatcttccccgaacgcctgcttaaaaccctgcaacagggtctgccagacttcaccagtcaaagcatgttgcaaaactttaggaacttta tcctagagcgttcaggaattctgcccgccacctgctgtgcgcttcctagcgactttgtgcccattaagtaccgtgaatgccctccgccgct ttggggtcactgctaccttntgcagctagccaactaccttgcctaccactccgacatcatggaagacgtgagcggtgacggcctactgg agtgtcactgtcgctgcaacctatgcaccccgcaccgctccctggtctgcaattcacaactgcttagcgaaagtcaaattatcggtacctt tgagctgcagggtccctcgcctgacgaaaagtccgcggctccggggttgaaactcactccggggctgtggacgtcggcttaccttcg caaatttgtacctgaggactaccacgcccacgagattaggttctacgaagaccaatcccgcccgccaaatgcggagcttaccgcctgc gtcattacccagggccacatccttggccaattgcaagccattaacaaagcccgccaagagtttctgctacgaaagggacggggggttt acttggacccccagtccggcgaggagctcaacccaatccccccgccgccgcagccctatcagcagccgcgggcccttgcttcccag gatggcacccaaaaagaagctgcagctgccgccgccgccacccacggacgaggaggaatactgggacagtcaggcagaggagg ttggacgaggaggaggagatgatggaagactgggacagcctagacgaggaagcttccgaggccgaagaggtgtcagacgaaac accgtcaccctcggtcgcattcccctcgccggcgccccagaaatcggcaaccgttcccagcattgctacaacctccgctcctcaggcg ccgccggcactgcccgttcgccgacccaaccgtagatgggacaccactggaaccagggccggtaagtctaagcagccgccgccgt tagcccaagagcaacaacagcgccaaggctaccgctcgtggcgcgtgcacaagaacgccatagttgcttgcttgcaagactgtggg ggcaacatctccttcgcccgccgctttcttctctaccatcacggcgtggccttcccccgtaacatcctgcattactaccgtcatctctacag cccctactgcaccggcggcagcggcagcaacagcagcggccacgcagaagcaaaggcgaccggatagcaagactctgacaaag cccaagaaatccacagcggcggcagcagcaggaggaggagcactgcgtctggcgcccaacgaacccgtatcgacccgcgagctt agaaacaggatttttcccactctgtatgctatatttcaacagagcaggggccaagaacaagagctgaaaataaaaaacaggtctctgcg ctccctcacccgcagctgcctgtatcacaaaagcgaagatcagcttcggcgcacgctggaagacgcggaggctctcttcagcaaata ctgcgcgctgactcttaaggactagtttcgcgccctttctcaaatttaagcgcgaaaactacgtcatctccagcggccacacccggcgc cagcacctgtcgtcagcgccattatgagcaaggaaattcccacgccctacatgtggagttaccagccacaaatgggacttgcggctgg agctgcccaagactactcaacccgaataaactacatgagcgcgggaccccacatgatatcccgggtcaacggaatccgcgcccacc gaaaccgaattctcctcgaacaggcggctattaccaccacacctcgtaataaccttaatccccgtagttggcccgctgccctggtgtacc aggaaagtcccgctcccaccactgtggtacttcccagagacgcccaggccgaagttcagatgactaactcaggggcgcagcttgcg 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tgtttctcaaccgctaaaaaaaacaaagtccaatataactttggaaacatccgcgccccttacagtcagctcaggcgccctaaccatggc cacaacttcgcctttggtggtctctgacaacactcttaccatgcaatcacaagcaccgctaaccgtgcaagactcaaaacttagcattgct accaaagagccacttacagtgttagatggaaaactggccctgcagacatcagcccccctctctgccactgataacaacgccctcactat cactgcctcacctcctcttactactgcaaatggtagtctggctgttaccatggaaaacccactttacaacaacaatggaaaacttgggctc aaaattggcggtcctttgcaagtggccaccgactcacatgcactaacactaggtactggtcagggggttgcagttcataacaatttgcta catacaaaagttacaggcgcaatagggtttgatacatctggcaacatggaacttaaaactggagatggcctctatgtggatagcgccgg tcctaaccaaaaactacatattaatctaaataccacaaaaggccttgcttttgacaacaccgcaataacaattaacgctggaaaagggttg gaatttgaaacagactcctcaaacggaaatcccataaaaacaaaaattggatcaggcatacaatataataccaatggagctatggttgc aaaacttggaacaggcctcagttttgacagctccggagccataacaatgggcagcataaacaatgacagacttactctttggacaacac cagacccatccccaaattgcagaattgcttcagataaagactgcaagctaactctggcgctaacaaaatgtggcagtcaaattttgggc actgtttcagctttggcagtatcaggtaatatggcctccatcaatggaactctaagcagtgtaaacttggttcttagatttgatgacaacgg agtgcttatgtcaaattcatcactggacaaacagtattggaactttagaaacggggactccactaacggtcaaccatacacttatgctgtt gggtttatgccaaacctaaaagcttacccaaaaactcaaagtaaaactgcaaaaagtaatattgttagccaggtgtatcttaatggtgaca agtctaaaccattgcattttactattacgctaaatggaacagatgaaaccaaccaagtaagcaaatactcaatatcattcagttggtcctgg aacagtggacaatacactaatgacaaatttgccaccaattcctataccttctcctacattgcccaggaataaagaatcgtgaacctgttgc atgttatgtttcaacgtgtttatttttcaattcgtattagtcatcgctattaccatggtgatgcggttttggcagtacatcaatgggcgtggatag cggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaat gtcgtaacaactccgccccattgacgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagagctggtttagtgaaccg tcagatccgctagagatccaccatgtttgtctttctcgtgctgctgcccctcgtgagcagccagtgcgtcaatctgacaacaaggaccca gctgccccccgcctacaccaactccttcacaagaggcgtgtattaccccgataaggtcttcagatccagcgtcctccacagcacccaa gatttgtttctgcctttcttcagcaacgtgacatggttccacgccattcatgtcagcggcacaaacggcacaaagaggtttgacaacccc gtgctccccttcaacgacggcgtgtacttcgccagcacagagaaatccaatatcattaggggctggatcttcggcacaacactggattc caagacccagtctctgctcattgtgaataacgccaccaacgtggtgattaaggtctgtgagtttcagttctgcaacgacccctttctggga gtctactaccacaagaataataagagctggatggagtccgagtttagggtgtacagctccgccaacaactgtaccttcgaatacgtgtc ccagcctttcctcatggatctggagggcaagcaaggcaatttcaaaaatctgagagagttcgtgttcaaaaacattgatggatacttcaa aatctacagcaagcatacccccattaatctggtgagggatctgccccaaggattctccgctctggaacctctggtggatctgcccattgg cattaacatcacaagattccagaccctcctcgccctccatagatcctatctgacccccggcgactcctccagcggatggacagccgga gctgccgcctactacgtgggctatctgcagccaagaacctttctgctgaagtacaacgagaacggcaccatcacagacgctgtcgatt gcgctctcgaccctctgagcgagaccaaatgcacactgaagagcttcaccgtggaaaagggcatctatcagaccagcaacttcagag tgcagcctaccgagagcattgtgaggtttcccaacatcaccaatctgtgtcctttcggcgaggtctttaatgccacaaggttcgcttccgt gtatgcttggaataggaagaggatcagcaattgcgtcgccgactattccgtcctctataacagcgcctccttctccaccttcaaatgttatg gcgtgtcccccaccaagctcaacgacctctgcttcaccaatgtgtacgctgactccttcgtcattaggggcgacgaggtgaggcaaatt gcccccggccagaccggcaagattgctgattacaactacaaactgcccgacgattttaccggctgcgtgatcgcttggaactccaaca atctggactccaaagtgggcggaaactacaattacctctacagactctttagaaaaagcaatctgaagcccttcgagagagacatctcc accgaaatctaccaagccggaagcacaccttgcaatggcgtcgagggatttaactgctacttccctctgcagagctacggctttcaacc taccaacggcgtcggatatcaaccctatagggtggtcgtgctgagctttgaactgctgcatgctcccgccaccgtctgcggacctaaga agagcaccaatctcgtcaaaaacaagtgcgtgaacttcaacttcaatggactgaccggcaccggcgtgctgaccgagagcaataaga agtttctgcccttccagcagttcggaagggatattgccgataccacagatgctgtgagggacccccaaaccctcgagattctggatatc accccttgcagcttcggaggagtgtccgtgatcacccccggaacaaacacctccaatcaagtggctgtgctgtaccaagacgtgaact gcacagaagtccccgtggccatccatgccgaccagctgacccctacatggagagtgtactccaccggcagcaatgtgttccagacaa gagccggatgcctcattggagctgaacacgtcaacaacagctacgagtgcgacattcccatcggcgccggcatttgtgcctcctatca gacccagaccaacagcccaagaagggctagaagcgtcgcttcccaatccatcattgcctacaccatgtctctgggagccgaaaactc cgtcgcctactccaacaatagcatcgccatccccaccaattttaccatctccgtgaccacagagattctgcccgtgtccatgacaaagac atccgtggactgcaccatgtacatctgtggcgacagcaccgagtgtagcaatctgctgctgcaatatggcagcttctgcacccagctga acagagccctcaccggcatcgccgtcgaacaagacaagaacacccaagaggtgttcgcccaagtgaagcaaatctacaagacccc ccctatcaaagatttcggaggattcaactttagccagattctgcccgatcctagcaagccttccaagaggagcttcatcgaggatctgct gtttaataaggtgacactggccgacgctggcttcattaaacagtacggcgattgtctgggcgacatcgctgctagggatctgatctgcg ctcagaagttcaacggactgacagtcctccctcctctgctgaccgacgagatgatcgctcagtataccagcgctctgctggctggaacc attaccagcggctggacattcggcgctggagccgccctccaaattccctttgccatgcagatggcctatagattcaacggcattggcgt cacccaaaatgtgctgtatgaaaatcagaagctgattgctaaccaattcaatagcgccattggcaagatccaagactctctgagctccac agccagcgccctcggaaagctgcaagacgtggtgaatcaaaacgcccaagctctgaacacactggtgaaacagctcagcagcaact ttggagccatcagcagcgtgctcaatgatatcctctctaggctggacaaagtggaggccgaagtccagatcgatagactcatcaccgg cagactccaatctctgcagacatacgtcacccaacagctcattagagctgccgaaatcagagcctccgccaatctggccgccaccaa gatgtccgagtgcgtgctgggacagagcaagagagtggacttctgtggcaagggataccatctgatgagcttcccccagagcgctcc ccatggagtggtctttctgcatgtcacatacgtgcccgcccaagagaagaacttcaccaccgctcccgccatttgccacgatggaaag gcccactttcccagagaaggagtgttcgtgagcaacggcacacactggtttgtcacccagagaaatttttacgagccccagattatcac caccgacaacaccttcgtgtccggaaactgcgatgtcgtgattggcatcgtgaacaacacagtctacgaccctctgcagcccgaactc gacagcttcaaggaagagctggacaagtacttcaagaatcacacatcccccgacgtggatctgggcgacattagcggcattaatgcct ccgtcgtcaacattcagaaggagattgatagactgaatgaagtcgccaagaacctcaatgagtctctgattgatctgcaagagctgggc aagtacgagcaatacatcaaatggccttggtacatctggctgggattcatcgctggactcatcgccatcgtgatggtcaccattatgctgt gttgcatgaccagctgctgcagctgtctgaagggctgctgcagctgcggaagctgctgcaagtttgacgaagacgactccgagcccg tgctgaagggcgtcaagctgcattatacataaactagtgctggaattcgcccttatagagtgctggaattcgcccttatagagtgctggaa ttcgcccttatatctagtaacggccgccagtgtgctggaattcgcccttataacttcgtatagcatacattatacgaagttattgttgacaatt aatcatcggcatagtatatcggcatagtataatacgacaaggtgaggaactaaaccatggccaagttgaccagtgccgttccggtgctc accgcgcgcgacgtcgccggagcggtcgagttctggaccgaccggctcgggttctcccgggacttcgtggaggacgacttcgccg gtgtggtccgggacgacgtgaccctgttcatcagcgcggtccaggaccaggtggtgccggacaacaccctggcctgggtgtgggtg cgcggcctggacgagctgtacgccgagtggtcggaggtcgtgtccacgaacttccgggacgcctccgggccggccatgaccgaga tcggcgagcagccgtgggggcgggagttcgccctgcgcgacccggccggcaactgcgtgcacttcgtggccgaggagcaggact gaataacttcgtatagcatacattatacgaagttataagggcgaattctgcagatatccatcctttaaaaaacctcccacacctccccctga acctgaaacataaaatgaatgcaattgttgttgttaacttgtttattgcagcttataatggttacaaataaagcaatagcatcacaaatttcac aaataaagcatttttttcactgcattctagttgtggtttgtccaaactcatcaatgtatcttaacaacgtgtttatttttcaattgcagaaagaatt gcagaaaatttcaagtcatttttcattcagtagtatagccccaccaccacatagcttatactaatcaccgtaccttaatcaaactcacagaa ccctagtattcaacctgccacctccctcccaacac...

Claims

CLAIMS1 . A method for inducing an immune response against a pathogen in a mammal, the method comprising administering a single cycle adenovirus (SC-Ad) comprising a nucleic acid sequence encoding an immunogen of said pathogen to said mammal, wherein said SC-Ad is administered by inhalation and infects a cell of said mammal, thereby causing expression of said immunogen leading to the induction of said immune response, without production of new infectious virions.

2. The method of claim 1, wherein said SC-Ad is formulated as a liquid formulation.

3. The method of claim 2, wherein the liquid formulation comprises NaCl, histidine, MgCl2, EDTA, polysorbate 80, sucrose, and ethanol.

4. The method of claim 1, wherein when said SC-Ad is formulated as a solid powder.

5. The method of any of claims 1 to 4, wherein said SC-Ad is administered in an aerosol.

6. The method of any of claims 1 to 5, wherein said SC-Ad is administered in a therapeutically effective amount of at least 1 x 1010SC-Ad viral particles.

7. The method of claim 6, wherein said SC-Ad is administered in a therapeutically effective amount of between 1 x 1010and 3 x 1011SC-Ad viral particles.

8. The method of any of claims 1 to 7, wherein the administration of said SC-Ad by inhalation is provided as a vaccine boost to a mammal that has received a previous vaccination to the pathogen before said subsequent administration of said SC-Ad by inhalation.

9. The method of claim 8, wherein the previous vaccination occurred at least one week, at least two weeks, at least three weeks, at least four weeks, at least five weeks, or at least six weeks before said subsequent administration of said SC-Ad by inhalation.

10. The method of claim 9, wherein the previous vaccination occurred at least 50 days, at least 60 days, at least 70 days, at least 80 days, at least 90 days, at least 100 days, at least 110days, at least 120 days, at least 130 days, at least 140 days, or at least 150 days before said subsequent administration of said SC-Ad by inhalation.

11. The method of any of claims 8 to 10, wherein said previous vaccination did not comprise administration of said SC-Ad.

12. The method of any of claims 8 to 10, wherein said previous vaccination comprised administration of said SC-Ad.

13. The method of any of claims 1 to 7, wherein the administration of said SC-Ad by inhalation is provided as a vaccine prime to a mammal that will receive a further vaccination boost to the pathogen after said administration of said SC-Ad by inhalation.

14. The method of claim 13, wherein said further vaccination will occur at least one week, at least two weeks, at least three weeks, at least four weeks, at least five weeks, or at least six weeks after said administration of said SC-Ad by inhalation.

15. The method of claim 14, wherein said further vaccination will occur at least 50 days, at least 60 days, at least 70 days, at least 80 days, at least 90 days, at least 100 days, at least 110 days, at least 120 days, at least 130 days, at least 140 days, or at least 150 days after said administration of said SC-Ad by inhalation.

16. The method of any of claims 13 to 15, wherein said further vaccination will not comprise administration of said SC-Ad.

17. The method of any of claims 13 to 16, wherein said further vaccination will comprise administration of said SC-Ad.

18. The method of any preceding claim, wherein said pathogen is a respiratory pathogen.

19. The method of claim 18, wherein the immunogen of said respiratory pathogen is selected from the group consisting of: a coronavirus immunogen (such as a SARS-CoV-2 immunogen, a SARS-CoV immunogen, a HCoVNL63 immunogen, a HKUl immunogen, or a MERS-CoV immunogen), an influenza immunogen, a respiratory syncytial virus (RSV)immunogen, a metapneumovirus (MPV) immunogen, a rhinovirus immunogen, a bocavirus immunogen, a parainfluenza virus (PIV) immunogen, a Streptococcus pneumoniae immunogen, a Bordetella pertussis immunogen, a Haemophilus influenzae immunogen, a Mycobacterium tuberculosis immunogen, a Klebsiella pneumonia immunogen; and an immunogen of a fungal respiratory pathogen.

20. The method of claim 19, wherein said pathogen is a coronavirus and the immunogen is a coronavirus immunogen comprising a coronavirus Spike polypeptide, or an immunogenic fragment thereof.

21. The method of any preceding claim, wherein a mucosal immune response is induced.

22. The method of claim 21 , wherein said mucosal immune response leads to an increase in the amount of serum IgG antibodies that bind to said immunogen.

23. The method of claim 21 or claim 22, wherein said mucosal immune response leads to an increase in the amount of IgA antibodies that bind to said immunogen.

24. The method of any preceding claim, wherein expression of said immunogen in said cell leads to an increase in the amount of serum IgA antibodies that bind to said immunogen.

25. The method of any preceding claim, wherein expression of said immunogen in said cell leads to an increase in the amount of secretory IgA antibodies that bind to said immunogen.

26. The method of any preceding claim, wherein expression of said immunogen in said cells leads to an increase in antibodies of a neutralising immune response.

27. The method of any preceding claim, wherein expression of said immunogen in said cells leads to an increased T cell response to the pathogen.

28. The method of claim 27, wherein expression of said immunogen in said cells leads to an increase in a systemic T cell response to the pathogen.

29. The method of claim 27 or claim 28, wherein expression of said immunogen in said cells leads to an increase in a mucosal T cell response to the pathogen.

30. The method of any preceding claim, wherein expression of said immunogen in said cells leads to an increase in a pro-inflammatory immune response.

31. The method of any preceding claim, wherein expression of said immunogen in said cells leads to an increase in expression of one, more or all of the following molecules: TNF-a, IFN-γ, IL-1, IL-6, IL-12, IL-18, and GM-CSF.

32. The method of any preceding claim, wherein said SC-Ad comprises a genome lacking at least a portion of a nucleic acid sequence that encodes an adenovirus polypeptide, and wherein said SC-Ad comprises said adenovirus polypeptide.

33. The method of claim 32, wherein said adenovirus polypeptide is selected from the group consisting of a fiber polypeptide, a V polypeptide, a hexon polypeptide, a penton base polypeptide, and a pllla polypeptide.

34. A single cycle adenovirus (SC-Ad) comprising a nucleic acid sequence encoding an immunogen of a pathogen for use in inducing an immune response against the pathogen in a mammal, wherein the SC-Ad is for administration by inhalation to infect a cell of the mammal thereby causing expression of the immunogen without production of new infectious virions, thus inducing the immune response.

35. The SC-Ad for use according to claim 34, wherein the SC-Ad is formulated as a liquid formulation.

36. The SC-Ad for use according to claim 35, wherein the liquid formulation comprises NaCl, histidine, MgCl2, EDTA, polysorbate 80, sucrose, and ethanol.

37. The SC-Ad for use according to claim 34, wherein when the SC-Ad is formulated as a solid powder.

38. The SC-Ad for use according to any one of claims 34 to 37, wherein the SC-Ad is for administration in an aerosol.

39. The SC-Ad for use according to any one of claims 34 to 38, wherein the SC-Ad is for administration in a therapeutically effective amount of at least 1 x 1010SC-Ad viral particles.

40. The SC-Ad for use according to any one of claims 34 to 39, wherein the SC-Ad is for administration in a therapeutically effective amount of between 1 x 1010and 3 x 1011SC-Ad viral particles.

41. The SC-Ad for use according to any one of claims 34 to 40, wherein the administration of the SC-Ad by inhalation is for use as a vaccine boost to a mammal that has previously received a previous vaccination to the pathogen before the subsequent administration of the SC-Ad by inhalation.

42. The SC-Ad for use according to claim 41, wherein the previous vaccination was received at least one week, at least two weeks, at least three weeks, at least four weeks, at least five weeks, or at least six weeks before said administration of the SC-Ad by inhalation.

43. The SC-Ad for use according to claim 42, wherein the previous vaccination was received at least 50 days, at least 60 days, at least 70 days, at least 80 days, at least 90 days, at least 100 days, at least 110 days, at least 120 days, at least 130 days, at least 140 days, or at least 150 days before said administration of the SC-Ad by inhalation.

44. The SC-Ad for use according to any of claims 41 to 43, wherein the previous vaccination did not comprise administration of the SC-Ad.

45. The SC-Ad for use according to any of claims 41 to 43, wherein the previous vaccination comprised administration of the SC-Ad.

46. The SC-Ad for use according to any of claims 34 to 40, wherein the administration of said SC-Ad by inhalation is for use as a vaccine prime to a mammal that will receive a further vaccination boost to the pathogen after said administration of said SC-Ad by inhalation.

47. The SC-Ad for use according to claim 46, wherein the further vaccination boost will be administered at least one week, at least two weeks, at least three weeks, at least four weeks, at least five weeks, or at least six weeks after said administration of the SC-Ad by inhalation.

48. The SC-Ad for use accordingto claim 47, wherein the further vaccination boost will be administered atleast 50 days, atleast 60 days, at least 70 days, at least 80 days, atleast 90 days, at least 100 days, at least 110 days, atleast 120 days, at least 130 days, at least 140 days, or at least 150 days after said administration of the SC-Ad by inhalation.

49. The SC-Ad foruse according to any of claims 46 to 48, wherein said further vaccination will not comprise administration of said SC-Ad.

50. The SC-Ad foruse according to any of claims 46 to 48, wherein said further vaccination will comprise administration of said SC-Ad.

51. The SC-Ad foruse according to any one of claims 34 to 50, wherein the pathogen is a respiratory pathogen.

52. The SC-Ad for use according to claim 51, wherein the immunogen of the respiratory pathogen is selected from the group consisting of: a coronavirus immunogen (such as a SARS- CoV-2 immunogen, a SARS-CoV immunogen, a HCoV NL63 immunogen, a HKU1 immunogen, or a MERS-CoV immunogen), an influenza immunogen, a respiratory syncytial virus (RSV) immunogen, a m etap neumo virus (MPV) immunogen, a rhinovirus immunogen, a bocavirus immunogen, a parainfluenza virus (PIV) immunogen, a Streptococcus pneumoniae immunogen, a Bordetella pertussis immunogen, a Haemophilus influenzae immunogen, a Mycobacterium tuberculosis immunogen, and a Klebsiella pneumonia immunogen.

53. The SC-Ad for use according to claim 52, wherein the pathogen is a coronavirus, and the immunogen is a coronavirus immunogen comprising a coronavirus Spike polypeptide, or an immunogenic fragment thereof.

54. The SC-Ad for use according to any one of claims 34 to 53 , wherein a mucosal immune response is induced.

55. The SC-Ad for use according to claim 54, wherein the mucosal immune response leads to an increase in the amount of serum IgG antibodies that bind to said immunogen.

56. The SC-Ad for use according to claim 54 or claim 55, wherein the mucosal immune response leads to an increase in the amount of IgA antibodies that bind to said immunogen.

57. The SC-Ad for use according to any one of claims 34 to 56, wherein expression of the immunogen in said cell leads to an increase in the amount of serum IgA antibodies that bind to said immunogen58. The SC-Ad for use according to any one of claims 34 to 57, wherein expression of the immunogen in said cell leads to an increase in the amount of secretory IgA antibodies that bind to said immunogen59. The SC-Ad for use according to any one of claims 34 to 58, wherein expression of said immunogen in said cells leads to an increase in antibodies of a neutralising immune response.

60. The SC-Ad for use according to any one of claims 34 to 59, wherein expression of said immunogen in said cells leads to an increased T cell response to the pathogen.

61. The SC-Ad for use according to claim 60, wherein expression of said immunogen in said cells leads to an increase in a systemic T cell response to the pathogen.

62. The SC-Ad for use according to claim 60 or claim 61, wherein expression of said immunogen in said cells leads to an increase in a mucosal T cell response to the pathogen.

63. The SC-Ad for use according to any one of claims 34 to 62, wherein expression of said immunogen in said cells leads to an increase in a pro-inflammatory immune response.

64. The SC-Ad for use according to any one of claims 34 to 63, wherein expression of said immunogen in said cells leads to an increase in expression of one, more or all of the following molecules: TNF-a, IFN-γ, IL-1, IL-6, IL-12, IL-18, and GM-CSF.

65. The SC-Ad for use according to any one of claims 34 to 64, wherein the SC-Ad comprises a genome lacking at least a portion of a nucleic acid sequence that encodes an adenovirus polypeptide, and wherein the SC-Ad comprises said adenovirus polypeptide.

66. The SC-Ad for use according to claim 65, wherein the adenovirus polypeptide is selected from the group consisting of a fiber polypeptide, a V polypeptide, a hexon polypeptide, a penton base polypeptide, and a pllla polypeptide.

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