Compositions Comprising Plasma and Immunoglobulins for Use in Treating or Preventing Infectious Diseases - Patent application

Abstract

The present invention relates to a composition for use in treating or preventing a respiratory infection in a subject, the composition comprising plasma and immunoglobulins against a causative agent of the respiratory infection, the immunoglobulins being obtained from a donor subject who has developed a humoral immune response against the causative agent of the respiratory infection, and the composition being administered intranasally, and optionally orally. The invention further relates to a nasal, nasopharyngeal, or oronasal spray device comprising such a composition, and a nasal, nasopharyngeal, or oronasal spray comprising plasma and immunoglobulins against the causative agent of the respiratory infection, the immunoglobulins being obtained from a donor subject who has developed a humoral immune response against the causative agent of the respiratory infection.

Description

FIELD OF THE INVENTION

[0001] The present invention relates to a pharmaceutical composition for treating or preventing infectious diseases, more preferably respiratory infections caused by respiratory viruses. Specific examples of such viruses include Coronaviridae, more specifically SARS-CoV, MERS-CoV, or SARS-CoV-2, which cause COVID-19. BACKGROUND OF THE INVENTION

[0002] Infectious disease epidemics pose a significant threat to public health. Passive immunotherapy, achieved by transferring specific antibodies from convalescent donors to patients, has been used to treat infectious diseases, including respiratory infections, for over a century and was awarded the first Nobel Prize in Medicine to Emil von Behring in 1901. However, despite this, the efficacy and safety of convalescent plasma have only been demonstrated in a limited number of case studies.

[0003] More recently, several multicenter, open-label, randomized clinical trials have been designed to demonstrate improved clinical outcomes after intravenous treatment with convalescent plasma (CCP) (containing high-titer anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies) for coronavirus disease 2019 (COVID-19) (a respiratory infection). However, despite over 10 published studies and over 100 ongoing studies, the clinical effectiveness of intravenous CCP for COVID-19 appears to be limited to a subset of immunocompromised patients. Results from completed clinical trials, such as the DAWn-PLASMA study in Belgium, the CAPSID study in Germany, and the RECOVERY study in the UK, have shown that intravenous administration of convalescent plasma in hospitalized patients with COVID-19 has little or no effect. Furthermore, intravenous transfusion of CCP into healthy individuals to prevent infection, such as SARS-CoV-2 infection, is problematic. First, it is logistically a nightmare because transfusions can only be performed in hospitals by authorized personnel under medical supervision. Second, there is always a risk (however minimal) of transfusion complications. Third, there is no evidence that transfusion of CCP benefits people. And finally, given that this plasma is not in infinite supply, not everyone can receive a transfusion, so ethical deontological concerns apply.

[0004] Over the past two decades, several betacoronaviruses (lineages B and C) have emerged, causing severe and potentially fatal respiratory illnesses. Their emergence in humans has primarily been via zoonotic transmission. The three betacoronaviruses that have caused the most clinically severe viral infections to date are the 2003 severe acute respiratory syndrome coronavirus (SARS-CoV), the 2013 Middle East respiratory syndrome coronavirus (MERS-CoV), and the 2019 severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The latter has caused a pandemic that has infected an estimated 253 million people and killed 5 million worldwide to date. Like all respiratory viruses, SARS-CoV-2 initiates infection in the upper respiratory tract. Furthermore, COVID-19 patients experience symptoms such as shortness of breath, fever, and dry cough. They may also develop a sudden, life-threatening inflammatory response (acute respiratory distress syndrome (ARDS)) in which fluid rapidly accumulates in the lungs, making oxygen absorption difficult.

[0005] Several vaccines are commercially available for respiratory infections, including COVID-19. While these vaccines generally prevent hospitalization, new variants continue to emerge that evade current humoral defenses and increase disease transmission. Additionally, older adults and patients with comorbidities receive limited or no vaccine protection. Furthermore, access to vaccines is limited or delayed in low- and middle-income countries. Vaccine hesitancy also creates gaps in population-wide protection.

[0006] Because the development of new antiviral drugs and vaccines is a time-consuming process, there is a need for new, readily available, and clinically relevant methods to treat or prevent infections with agents that cause respiratory infections in general, and in particular infections with pathogens transmitted via the nasopharyngeal route, whether causing local infection and / or more systemic effects. Summary of the Invention

[0007] As evidenced in the Examples section illustrating certain representative embodiments of the present invention, the inventors have discovered that infection and transmission of a causative agent (i.e., a pathogen such as a virus, bacterium, bacterial spore, or fungus) that enters and is transmitted via the nasopharyngeal route can be prevented or inhibited by nasopharyngeal administration (i.e., ingestion via the nasal or oropharyngeal mucosa) to a subject to be treated of a composition comprising plasma and immunoglobulin against the causative agent, wherein the immunoglobulin is obtained from a donor subject that has developed a humoral immune response against the causative agent.

[0008] More specifically, the inventors have discovered that respiratory infections, e.g., respiratory infections caused by a causative agent (i.e., a pathogen such as a virus, bacterium, bacterial spore, or fungus), preferably a virus, can be treated and / or prevented by nasopharyngeal administration (i.e., ingestion via the nasal or oropharyngeal mucosa) to a subject of a composition comprising plasma and immunoglobulins against the causative agent of the respiratory infection, wherein the immunoglobulins are obtained from a donor subject that has developed a humoral immune response against the causative agent of the respiratory infection. This results in a rapid-response product that can help protect the population from widespread infection by respiratory pathogens, such as respiratory viruses, before traditional vaccines, antibiotics, or antivirals are developed against those pathogens.

[0009] As proof of concept (see the Examples section), we demonstrated in a SARS-CoV-2 hamster transmission model that intranasal administration of (i) convalescent plasma obtained from a human donor subject who had recovered from SARS-CoV-2 infection or (ii) convalescent plasma obtained from a human donor subject who had recovered from SARS-CoV-2 infection and been vaccinated against SARS-CoV-2 to hamsters one day before and four days during exposure (i.e., a total of five consecutive days) to SARS-CoV-2-infected hamsters significantly reduced viral RNA and intact infectious virus in the lungs of the hamsters compared to control hamsters. Interestingly, administration of the convalescent plasma to the hamsters resulted in a significant reduction in intact infectious virus in the lungs of the hamsters compared to administration of purified human immunoglobulin in the absence of plasma. Furthermore, administration of non-immune hamster plasma containing purified human immunoglobulins against the causative agent of the respiratory infection resulted in a significant reduction of viral genetic material (RNA) in the lungs of the hamsters compared to administration of purified human immunoglobulins in the absence of non-immune hamster plasma, where the immunoglobulins were obtained from a donor subject that had developed a humoral immune response against the causative agent of the respiratory infection.

[0010] Similar to the data on administration of human convalescent plasma, we also demonstrated in a SARS-CoV-2 hamster transmission model that intranasal administration of convalescent plasma collected from hamster donor subjects who had recovered from SARS-CoV-2 infection to hamsters one day before and for four days during exposure (i.e., a total of five consecutive days) to SARS-CoV-2-infected hamsters significantly reduced viral RNA and infectious virus in the lungs of the hamsters compared to control hamsters. We also showed that non-immunized hamster plasma supplemented with purified human immunoglobulins was also able to significantly reduce viral RNA and intact infectious virus in the lungs of infected hamsters compared to control hamsters. Without wishing to be bound by theory, the inventors believe that the observed effects are due, at least in part, to (i) favorable interactions between plasma components such as coagulation factors, complement cascade factors, cytokines and chemokines and antibodies against the causative agents of the respiratory infection, and / or (ii) a favorable effect of the plasma matrix viscosity on the availability of immunoglobulins.

[0011] Thus, the present invention generally provides a method for preventing infection and transmission of a causative agent, such as a virus, bacterium, bacterial spore, or fungus, that enters and is transmitted via the nasopharyngeal route, by nasopharyngeal administration (i.e., ingestion via the nasal or oropharyngeal mucosa) to a subject to be treated of a composition comprising plasma components and immunoglobulin against the causative agent, wherein the immunoglobulin is obtained from a donor subject that has developed a humoral immune response against the causative agent. Disease caused by the causative agent may be localized (i.e., to the nasopharyngeal region), directed generally to the lower respiratory system (i.e., including the lungs), systemic (i.e., the nasopharyngeal region is the only portal for the causative agent to enter the subject's bloodstream, causing systemic disease), or directed to the gastrointestinal system.

[0012] Therefore, more specifically, the present invention provides the following aspects. Aspect 1 A method for preventing infection and transmission of a causative agent, such as a virus, bacterium, bacterial spore, or fungus, which enters and is transmitted via the nasopharyngeal route, by nasopharyngeal administration (i.e., ingestion via the nasal or oropharyngeal mucosa) to a subject to be treated of a composition comprising plasma and immunoglobulin against the causative agent, wherein the immunoglobulin is obtained from a donor subject who has developed a humoral immune response against the causative agent. The present invention also provides a composition for use in preventing infection and transmission of the causative agent as defined herein. In any of the embodiments herein, the plasma can be in the form of (whole plasma, preferably convalescent) plasma or an isolated component thereof. Aspect 2 A method for preventing infection and transmission of a causative agent of a respiratory infection, comprising nasopharyngeal administration (i.e., ingestion via the mucous membranes of the nose or oropharynx) of a composition comprising plasma and immunoglobulins against said causative agent, wherein said immunoglobulins are obtained from a donor subject who has developed a humoral immune response against said causative agent. The above aspects also provide a composition for use in preventing infection and transmission of a causative agent that causes a respiratory infection, and for treating or preventing a respiratory infection in a subject, wherein: the composition comprises plasma and immunoglobulins against a causative agent of the respiratory infection, the immunoglobulins being obtained from a donor subject who has developed a humoral immune response against the causative agent of the respiratory infection; and The composition is administered by nasopharyngeal administration (ie, ingestion through the mucous membranes of the nose or oropharynx). Aspect 3 3. A method or composition for use according to embodiment 1 or 2 for use in reducing the progression of an infection, such as an upper respiratory tract infection, to the lower respiratory tract, systemic or gastrointestinal system and / or preventing the spread of said infection in a population. Aspect 4 4. A method or composition for use according to any one of aspects 1 to 3, wherein said composition comprises at least 70% (v / v) of said plasma and immunoglobulins against said causative agent, wherein said immunoglobulins are obtained from a donor subject that has developed a humoral immune response against said causative agent. Aspect 5 5. The method or composition for use according to any one of aspects 1 to 4, wherein said plasma is not rich in platelets, preferably said plasma is not platelet rich plasma (PRP). Aspect 6 5. The method or composition for use according to any one of aspects 1 to 4, wherein said composition comprises convalescent plasma obtained from a donor subject that has developed a humoral immune response to said causative agent. Aspect 7 The composition comprises: - before the subject is exposed to the causative agent (i.e., to avoid transmission to unexposed subjects); - before the subject becomes infected with the causative agent (i.e., to avoid transmission to exposed but uninfected subjects); - after the subject has been exposed to the causative agent (i.e., to avoid infection and development of disease); or - after the subject has been infected with the causative agent (i.e., to prevent the onset and / or progression of disease) A method or composition for use according to any one of aspects 1 to 6, administered to said subject. Aspect 8 A composition for use according to any one of aspects 1 to 6, wherein the causative agent of a respiratory infection is a virus, bacterium, bacterial spore, or fungus, preferably a virus. Aspect 9 9. The method or composition for use according to any one of aspects 1 to 8, wherein the disease is a respiratory infection, more preferably a viral respiratory infection, and more preferably the causative agent of the viral respiratory infection is selected from the group consisting of Coronaviridae, Respiratory Syncytial Virus (RSV), Influenza virus, Parainfluenza virus, Metapneumovirus, Rhinovirus, Adenovirus, and Bocavirus, and preferably the respiratory virus is a Coronaviridae virus. Aspect 10 10. The method or composition for use according to embodiment 9, wherein said Coronaviridae virus is a severe acute respiratory syndrome-associated coronavirus, most preferably severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2; causing COVID-19). Aspect 11 11. The method or composition for use according to any one of aspects 1 to 10, wherein the composition is a pulmonary aerosol formulation, nasal drops, oral nasal drops, nasal spray, oral nasal spray, or nasopharyngeal drops or spray. Aspect 12 12. The method or composition for use according to any one of aspects 1 to 11, wherein said composition is used as an adjunct to conventional (respiratory) infectious disease treatment, prevention or vaccination methods. Aspect 13 12. The method or composition for use according to any one of aspects 1 to 11, wherein said composition is used in the absence of conventional methods for treating, preventing or vaccinating against (respiratory) infectious diseases. Aspect 14 A nasal, nasopharyngeal or oronasal spray comprising plasma and immunoglobulins against a causative agent of a (respiratory) infection as defined herein, wherein said immunoglobulins are obtained from a donor subject who has developed a humoral immune response against said causative agent. Preferably, said spray comprises a composition as defined in any one of aspects 1 to 13. Aspect 15 A nasal, nasopharyngeal or oronasal spray device comprising plasma and a composition comprising immunoglobulins against a causative agent of a (respiratory) infection as defined herein, wherein said immunoglobulins are obtained from a donor subject who has developed a humoral immune response against said causative agent. Preferably, said spray comprises a composition as defined in any one of aspects 1 to 13. Aspect 16 A method of treating or preventing a respiratory infection in a subject, comprising administering to the subject a composition comprising plasma and immunoglobulins against a causative agent of the respiratory infection, wherein the immunoglobulins are obtained from a donor subject that has developed a humoral immune response against the causative agent of the respiratory infection; The composition is administered intranasally or to the oropharyngeal region (ie, uptake via the nasal or oropharyngeal mucosa). Aspect 17 20. The method of embodiment 16, for use in reducing the progression of an upper respiratory tract infection to the lower respiratory tract and / or for use in preventing the transmission of an upper respiratory tract infection in a population. Aspect 18 18. The method of aspect 16 or 17, wherein the composition comprises at least 70% (v / v) of the plasma and immunoglobulins against the causative agent of the respiratory infection, wherein the immunoglobulins are obtained from a donor subject that has developed a humoral immune response against the causative agent of the respiratory infection. Aspect 19 Aspect 19. The method according to any one of aspects 16 to 18, wherein the plasma is not rich in platelets, preferably the plasma is not platelet-rich plasma (PRP). Aspect 20 20. The method of any one of aspects 16-19, wherein the composition comprises convalescent plasma obtained from a donor subject that has developed a humoral immune response against a causative agent of the respiratory infection. Aspect 21 The composition comprises: - before the subject is exposed to the agent causing the respiratory infection (i.e., to avoid transmission to unexposed subjects); - before the subject becomes infected with the agent causing the respiratory infection (i.e., to avoid transmission to exposed but uninfected subjects); - after the subject has been exposed to the agent causing the respiratory infection (i.e., to avoid infection and development of the respiratory disease); or - after the subject has been infected with the agent causing the respiratory infection (i.e., to prevent the onset and / or progression of respiratory disease). 21. The method according to any one of aspects 16 to 20, wherein the method is administered to the subject. Aspect 22 Aspect 22. The method according to any one of aspects 16 to 21, wherein the causative agent of the respiratory infection is a virus, bacterium, bacterial spore, or fungus, preferably a virus. Aspect 23 23. The method of any one of Aspects 16 to 22, wherein the respiratory infection is a viral respiratory infection and the causative agent of the viral respiratory infection is selected from the group consisting of a Coronaviridae virus, a respiratory syncytial virus (RSV), an influenza virus, a parainfluenza virus, a metapneumovirus, a rhinovirus, an adenovirus, and a bocavirus, and preferably the respiratory virus is a Coronaviridae virus. Aspect 24 24. The method of aspect 23, wherein the Coronaviridae virus is a severe acute respiratory syndrome-associated coronavirus, most preferably severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2; causing COVID-19). Aspect 25 Aspect 25. The method of any one of aspects 16 to 24, wherein the composition is a pulmonary aerosol formulation, nasal drops, oral nasal drops, nasal spray, oral nasal spray, or nasopharyngeal drops or spray. Aspect 26 26. The method of any one of aspects 16 to 25, wherein the composition is used as an adjunct to conventional respiratory infection treatment, prophylaxis or vaccination methods. Aspect 27 Aspect 27. The method of any one of aspects 16 to 26, wherein the composition is used in the absence of conventional methods for treating, preventing, or vaccinating against respiratory infections. Aspect 28 Use of a composition comprising plasma and immunoglobulins against a causative agent of an infectious disease as defined herein in the manufacture of a medicament for the treatment and / or prevention of said infectious disease in a subject, wherein said immunoglobulins are obtained from a donor subject who has developed a humoral immune response against the causative agent of said infectious disease; The composition is administered intranasally or to the oropharyngeal region (ie, uptake via the nasal or oropharyngeal mucosa). Aspect 29 29. Use according to embodiment 28 for reducing the progression of an upper respiratory tract infection to the lower respiratory tract, systemic or gastrointestinal system and / or for preventing the spread of said infection in a population. Aspect 30 30. The use of aspect 28 or 29, wherein the composition comprises at least 70% (v / v) of the plasma and immunoglobulins against the causative agent of the infectious disease, wherein the immunoglobulins are obtained from a donor subject that has developed a humoral immune response against the causative agent of the infectious disease. Aspect 31 The use according to any one of aspects 28 to 30, wherein the plasma is not rich in platelets, preferably the plasma is not platelet-rich plasma (PRP). Aspect 32 32. The use of any one of aspects 28 to 31, wherein the composition comprises convalescent plasma obtained from a donor subject that has developed a humoral immune response against the causative agent of the infectious disease. Aspect 33 The composition comprises: - before the subject is exposed to the agent causing the infectious disease (i.e., to avoid transmission to unexposed subjects); - before the subject becomes infected with the causative agent of the infectious disease (i.e., to avoid transmission to exposed but uninfected subjects); - after the subject has been exposed to the agent causing the infectious disease (i.e., to avoid infection and development of respiratory disease); or - after the subject has been infected with the causative agent of the infectious disease (i.e., to avoid the onset and / or progression of respiratory disease) 33. The use according to any one of aspects 28 to 32, wherein the use is administered to said subject. Aspect 34 34. The use according to any one of aspects 28 to 33, wherein the causative agent of the infectious disease is a virus, bacterium, bacterial spore, or fungus, preferably a virus. Aspect 35 35. The use according to any one of aspects 28 to 34, wherein the infection is a respiratory infection, such as a viral respiratory infection, and more preferably the causative agent of the viral respiratory infection is selected from the group consisting of a coronavirus, a respiratory syncytial virus (RSV), an influenza virus, a parainfluenza virus, a metapneumovirus, a rhinovirus, an adenovirus, and a bocavirus, and preferably the respiratory virus is a coronavirus. Aspect 36 36. The use according to aspect 35, wherein the Coronaviridae virus is a severe acute respiratory syndrome-associated coronavirus, most preferably severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2; which causes COVID-19). Aspect 37 37. The use according to any one of aspects 28 to 36, wherein the composition is a pulmonary aerosol formulation, nasal drops, oral nasal drops, nasal spray, oral nasal spray, or nasopharyngeal drops or spray. Aspect 38 38. The use according to any one of aspects 28 to 37, wherein the composition is used as an adjunct to conventional infectious disease treatment, prophylaxis or vaccination methods. Aspect 39 39. The use according to any one of aspects 28 to 38, wherein the composition is used in the absence of conventional methods for treating, preventing or vaccinating against infectious diseases. Aspect 40 14. The method or composition for use according to any one of aspects 1 to 13, wherein said composition induces a humoral immune response against said causative agent in a subject to which said composition is administered. Aspect 41 A method for inducing a humoral immune response against a pathogen invading or transmitting in the nasopharyngeal region of a subject in need thereof, comprising administering to the subject a composition comprising plasma and immunoglobulins against the pathogen, wherein the immunoglobulins are obtained from a donor subject that has developed a humoral immune response against the pathogen, and wherein the composition is administered intranasally or to the nasopharyngeal region. Preferably, the composition is as defined in any one of aspects 1 to 13. Aspect 42 A method for producing a vaccine comprising combining plasma and a composition comprising immunoglobulins against a pathogen that invades or spreads in the nasopharyngeal region with one or more pharmaceutically acceptable excipients, wherein said immunoglobulins are obtained from a donor subject that has developed a humoral immune response against said pathogen. Preferably, said composition is as defined in any one of aspects 1 to 13.

[0013] These and further aspects of the invention, as well as preferred embodiments, are described in the following sections and in the appended claims, the subject matter of which is specifically incorporated herein. [Brief explanation of the drawings]

[0014] Figure 1. Intranasal administration of human (convalescent) plasma in a SARS-CoV-2 hamster infection model. Sentinel hamsters were given multiple human plasma derivatives, namely vaccinated COVID-19 convalescent plasma (VCCP), COVID-19 convalescent plasma (CCP), human antibodies purified from VCCP (purified hIg), or non-immune human plasma (NIP) donated before the COVID-19 pandemic. ヒト) was administered intranasally. Phosphate-buffered saline was included as a negative buffer control. The levels of (a-c) (IgG (a), IgA (b), and IgM (c)) binding to the SARS-CoV-2 receptor-binding domain (RBD) in human plasma samples were measured by isotype ELISA and calibrated to the international WHO standard (BAU / mL) (n = 3, each tested in duplicate). The concentrations of IgG (e) and IgA (f) in human plasma were measured by ELISA (μg / mL) (n = 3, each tested in duplicate); (g) Neutralizing capacity (expressed in IU / mL) of plasma samples tested by inhibition ELISA (n = 3, each tested in duplicate); (h) Inhibition of plaque formation by (convalescent) plasma. Plaque formation was tested in VeroE6 cultures using the SARS-CoV-2 Wuhan strain (n = 1, tested in duplicate); (i) Preparation of Syrian hamsters for transmission tests. Sentinel hamsters were treated daily with human plasma or buffer (25 μL per nostril). On day 0, index hamsters were given 2 x 10 6 TCID 50 The index hamster was killed on day 4 post-infection, and the sentinel hamster was killed on day 5 (VCCP, purified hIg, and NIP). ヒト (j) Viral RNA levels in the lungs of hamsters on day 4 (index) or day 5 (sentinel) as quantified by RT-qPCR and expressed as RNA copies / mg lung tissue; (k) log 10 TCID 50Infectious viral load in the lungs of index hamsters and hamsters prophylactically treated with various plasma samples, expressed as / mg lung tissue; (l) Cumulative lung scores from H&E-stained sections of the lungs of infected (index) and infected, prophylactically treated (sentinel) hamsters; (d) H&E images of the lungs of sentinel hamsters, comparing buffer-treated and CCP-treated hamsters on days 4 and 5. Pneumocyte hyperplasia was observed in the lungs of CCP-treated hamsters (blue box). Scale bar, 200 μm. Median values ​​with individual data and 95% CIs are shown. Statistical significance between groups was calculated by Kruskal-Wallis with Dunn's post-hoc test. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001; ns=not significant. IU / mL = International Units / milliliter. BAU / mL = Bound Antibody Units / milliliter.

[0015] Figure 2. Intranasal administration of hamster plasma in a SARS-CoV-2 hamster infection model Sentinel hamsters were given several hamster plasma derivatives, namely COVID-19 convalescent hamster plasma (CCP). ハムスター ), purified human antibodies from VCCP spiked into non-immune hamster plasma (NIP ハムスター )(NIP ハムスター purified hIg in IgG, or NIP ハムスター (a) Preparation of Syrian hamster infection test. Sentinel hamsters were treated daily with hamster plasma (25 μL / nostril). On day 0, index hamsters were given 2x10 6 TCID 50(b) Neutralization capacity (expressed in international WHO standard units (IU / mL)) of plasma samples tested by inhibition ELISA (n=3, each tested in duplicate); (c) Weight change of index hamsters at 4 days post-infection (pi) and sentinel hamsters at 5 days post-infection (pi) shown as percentage and normalized to body weight on day 0 of infection / treatment (n=6 for all cohorts except for buffer control n=14 from two independent experiments); (d) Viral RNA levels in the lungs of hamsters at 4 days post-infection or pi (index) or 5 days post-infection (sentinel) expressed as RNA copies / mg lung tissue; (e) log 10 TCID 50 Infectious viral load in the lungs of the index hamster and hamsters treated prophylactically with various hamster plasma samples on day 4 or pi, expressed as / mg lung tissue; (f) Cumulative lung scores from H&E stained sections of the lungs of infected hamsters (index) and exposed and prophylactically treated hamsters (sentinel); (g) H&E images of the lungs of hamsters treated with plasma on day 5. CCP ハムスターR No inflammation or pneumocyte hyperplasia was observed in the lungs of hamsters treated with IFN-α. Scale bar, 200 μm. Median individual data and 95% CI are shown. Statistical significance between groups was calculated by Kruskal-Wallis with Dunn's post-hoc test. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ns = not significant. IU / mL = international units per milliliter.

[0016] Figure 3. Plaque reduction neutralization assay with delta (a) and omicron (b) SARS-CoV-2 mutants. VCCP, CCP and NIP ヒト Serial dilutions of 1000 ng / ml were tested for their ability to inhibit plaque formation in Vero E6 cells upon infection with SARS-CoV-2 mutants. 10 The % inhibition is shown for each plasma dilution of CCP expressed as log (%) (n=1, each tested in duplicate).

[0017] Figure 4. Intranasal administration of human plasma in a SARS-CoV-2 hamster infection model Syrian hamsters were given low-titer COVID-19 convalescent plasma (CCP) 低 (a) Syrian hamster infection test preparation. Hamsters were treated with CCP, mAb, or buffer control (25 μL / nostril) on day 1 and 10 μL / nostril on day 0. 3 TCID 50 The second treatment was with the Wuhan strain of HIV 1 one hour before intranasal inoculation; (b-d) Levels of IgG (b), IgA (c), and IgM (d) in human plasma samples were measured by isotype ELISA and calibrated to the international WHO standard (BAU / mL) (n=3, each tested in duplicate). (f) and IgA (g) concentrations in human plasma were measured by ELISA (μg / mL) (n=3, each tested in duplicate); (h) Neutralization capacity of plasma samples tested by inhibition ELISA and expressed in international WHO standard units (IU / mL) (n=3, each tested in duplicate); (e) Plaque reduction neutralization test (PRNT) with serial dilutions of mAbs against Vero E6 cells infected with the SARS-CoV-2 Wuhan strain, expressed as plaque formation (%) normalized to uninfected cells (n=1, each tested in duplicate); (i) Body weight change at 4 days post-infection (pi) shown as a percentage and normalized to body weight at day 0 of infection / treatment (n=6); (j) Viral RNA levels in hamster lungs at 4 days post-infection (pi) expressed as RNA copies / mg lung tissue; (k) log 10 TCID 50(l) Infectious viral load in the lungs on day 4 (pi) of hamsters prophylactically treated with CCP or buffer on days 1 and 0, expressed as / mg lung tissue; (l) Cumulative lung scores from H&E-stained sections of lungs from infected and prophylactically treated hamsters. Individual data and median values ​​with 95% CI are shown. Statistical significance between groups was calculated by Kruskal-Wallis with Dunn's post-hoc test. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001; ns = not significant. IU / mL = International Units / milliliter. BAU / mL = Bound Antibody Units / milliliter.

[0018] Figure 5. Intranasal administration of human and hamster plasma to naive Syrian hamsters Naive Syrian hamsters were injected with vaccinated COVID-19 convalescent plasma or serum (VCCP or VCCS), purified human antibodies from VCCP spiked in buffer (purified hIg in buffer), and purified human antibodies from VCCP spiked in non-immunized hamster plasma (NIP). ハムスター )(NIP ハムスター purified hIg from plasma, non-immune human plasma (NIP) donated before the COVID-19 pandemic ヒト ), NIP ハムスター , and NIS ハムスター (a) Syrian hamster safety test preparation. Unvaccinated hamsters were treated with VCCP, VCCS, NIP, or a buffer control. ハムスター , NIS ハムスター , NIP ヒト , purified hIg in buffer, NIP ハムスター (b) Cumulative lung scores from H&E-stained sections of lungs from hamsters treated with plasma / serum or buffer (n=6); (c) H&E images of lungs from buffer- and CCP-treated hamsters on day 5. NIP ヒトAlveolar epithelial cell hyperplasia was observed in the lungs of hamsters treated with IFN-γ (blue box). Scale bar, 200 μm. Median individual data and 95% CI are shown. Statistical significance between groups was calculated by Kruskal-Wallis with Dunn's post-hoc test. *p<0.05, ns = not significant.

[0019] Figure 6. Humoral immunity after intranasal administration of convalescent plasma: Experimental setup Abbreviations: CCP: convalescent plasma; TCID: tissue culture infectious dose.

[0020] As used herein, the singular forms "a," "an," and "the" include both singular and plural referents unless the context clearly dictates otherwise.

[0021] As used herein, the terms "comprising," "including," and "consisting of" are synonymous with "including," "including," or "containing," and are not inclusive or limiting and do not exclude additional, unrecited members, elements, or method steps. These terms also include "consisting of" and "consisting essentially of," which have well-established meanings in patent language.

[0022] The recitation of numerical ranges by endpoints includes each range and all values ​​and fractions subsumed within the recited endpoints.

[0023] As used herein, the terms "about" or "approximately" to refer to a measurable value such as a parameter, amount, duration, etc., are meant to encompass variations from the specified value, e.g., variations of no more than ±10%, preferably no more than ±5%, more preferably no more than ±1%, and even more preferably no more than ±0.1% of the specified value, provided that such variations are appropriate for the disclosed invention. It is to be understood that the value referred to by the modifier "about" is itself specifically, and preferably, disclosed.

[0024] The term "one or more" or "at least one," e.g., one or more members, or at least one member of a group of members, is self-explanatory, but by way of further example, the term includes reference to, among other things, any one of the aforementioned members, or any two or more of the aforementioned members, e.g., three or more, four or more, five or more, six or more, or seven or more of the aforementioned members, etc., and all of the aforementioned members. As another example, "one or more" or "at least one" may refer to 1, 2, 3, 4, 5, 6, 7, or more.

[0025] The discussion of the background of the invention herein is included to explain the context of the invention and is not an admission that any of the material mentioned was published, known, or part of common general knowledge in any country as of the priority date of any claim.

[0026] Throughout this disclosure, various publications, patents, and published patent specifications are referenced by an identifying citation. All documents cited herein are incorporated by reference in their entirety. In particular, the teachings or sections of documents specifically referenced herein are incorporated by reference.

[0027] Unless otherwise defined, all terms used in disclosing the present invention, including technical and scientific terms, have the meanings commonly understood by one of ordinary skill in the art to which this invention belongs. For further guidance, term definitions are included to better understand the teachings of the present invention. When a particular term is defined in connection with a particular aspect of the invention or a particular embodiment of the invention, such meaning shall also apply throughout the specification, i.e., in the context of other aspects or embodiments of the invention, unless otherwise defined.

[0028] In the following text, various aspects or embodiments of the present invention are defined in more detail. Each aspect or embodiment so defined may be combined with other aspects or embodiments, unless expressly indicated to the contrary. In particular, any feature indicated as being preferred or advantageous may be combined with any other feature indicated as being preferred or advantageous.

[0029] References throughout this specification to "one embodiment" or "an embodiment" mean that a particular feature, structure, or characteristic described in connection with an embodiment is included in at least one embodiment of the present invention. Thus, appearances of the phrase "one embodiment" or "an embodiment" in various places throughout this specification do not necessarily all refer to the same embodiment, although they may. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments, as would be apparent to one of ordinary skill in the art from this disclosure. Furthermore, while some embodiments described herein include some features but not others included in other embodiments, combinations of features from different embodiments are intended to form different embodiments within the scope of the present invention and as would be understood by one of ordinary skill in the art. For example, in the appended claims, any of the claimed embodiments may be used in any combination.

[0030] Surprisingly, and as supported in the Examples section illustrating certain exemplary embodiments of the present invention, the inventors have found that infections that originate in the nasopharyngeal region, more preferably respiratory infections such as COVID-19, can be treated and / or prevented by intranasal or nasopharyngeal administration of a composition comprising plasma components, preferably whole plasma, and immunoglobulins against the causative agent of said infection, wherein said immunoglobulins are obtained from a donor subject that has developed a humoral immune response against the causative agent of said infection.

[0031] As proof of concept (see the Examples section) and as described above, we conducted experiments in a SARS-CoV-2 hamster transmission and infection model. Hamsters are obligate nasal breathers and are frequently used to investigate human respiratory virus-induced diseases and evaluate the efficacy of vaccines or drugs. For SARS-CoV-2 infection, the hamster model we used is highly suitable because these animals express the angiotensin-converting enzyme 2 (ACE) receptor, a SARS-CoV-2 entry receptor, in their airways. Furthermore, intranasal SARS-CoV-2 inoculation of hamsters effectively induces viral replication in the upper respiratory tract, making this model well suited for preclinical testing.

[0032] Intranasal (e.g., nasal inhalation) or nasopharyngeal (e.g., via nasal or oral inhalation) administration of compositions containing plasma and immunoglobulins against the causative agent of an infection that begins in the nasopharyngeal region, more preferably a respiratory infection, may be applied, for example, as a spray, requiring the use of smaller amounts of plasma compared to intravenous administration of convalescent plasma, where the immunoglobulins are obtained from a donor subject who has developed a humoral immune response against the causative agent of the infection. Furthermore, plasma, such as convalescent plasma, is readily available, even in the very early stages of an epidemic, and is inexpensive, especially compared to recombinant (monoclonal) antibody or nanobody cocktails and small (antiviral) molecules. Intranasal or nasopharyngeal delivery of the plasma and immunoglobulin-containing compositions described herein, e.g., intranasal or nasopharyngeal delivery of convalescent plasma, may be used in the early stages of a pandemic for patients who are only partially or not protected at all by vaccination (e.g., many elderly patients, patients with underlying conditions that affect humoral immunity, etc.), and for patients living in low- and middle-income countries that suffer from limited or delayed access to vaccines.

[0033] Thus, intranasal or nasopharyngeal delivery of the plasma and immunoglobulin-containing compositions described herein, e.g., delivery of convalescent plasma, can reduce the burden on healthcare providers such as hospitals, nursing homes, and nursing homes, as well as primary care physicians and (outpatient) care providers, thereby reducing healthcare costs.

[0034] Accordingly, a first aspect provides a composition for use in the treatment and / or prevention of an infection originating in the nasopharyngeal region, more preferably a respiratory infection, in a subject, comprising: wherein the composition comprises plasma and immunoglobulins against the causative agent of the infectious disease, the immunoglobulins being obtained from a donor subject who has developed a humoral immune response against the causative agent of the infectious disease; and The compositions are administered intranasally or to the nasopharyngeal region.

[0035] A further aspect provides a method for treating and / or preventing an infection originating in the nasopharyngeal region, more preferably a respiratory infection, in a subject, comprising administering to said subject an effective amount of a composition comprising plasma and immunoglobulins against a causative agent of the infection, said immunoglobulins being obtained from a donor subject that has developed a humoral immune response against the causative agent of the infection, and wherein administration of said composition is intranasal or in the nasopharyngeal region.

[0036] In certain embodiments, the compositions comprising plasma and immunoglobulins described herein induce a humoral immune response against the causative agent in a subject to which the composition is administered.

[0037] In certain embodiments, the compositions comprising plasma and immunoglobulin described herein are compositions that stimulate a humoral immune response, hi certain embodiments, the compositions comprising plasma and immunoglobulin described herein are compositions for vaccination, such as first-line vaccines or vaccine-like compositions.

[0038] Thus, a further aspect provides a method of inducing a humoral immune response against a pathogen invading or transmitting in the nasopharyngeal region of a subject in need thereof, the method comprising administering to the subject a composition comprising plasma and immunoglobulin against the pathogen, the immunoglobulin being obtained from a donor subject that has developed a humoral immune response against the pathogen, and the composition being administered intranasally or to the nasopharyngeal region.

[0039] As used herein, the term "preventing" or "prevention" refers to a reduction in the risk of developing a disease or disorder (i.e., preventing the development of at least one clinical symptom of a disease in a subject, particularly a human subject, who may be exposed to or susceptible to the disease but who has not yet experienced or exhibited symptoms of the disease). In this context, the term prevention also encompasses reducing or preventing transmission of a causative agent and / or reducing or preventing infection by a causative agent, thereby actively preventing infection and / or the onset and progression of the disease.

[0040] The term "therapy" or "treatment" of any disease or disorder, in one embodiment, includes ameliorating the disease or disorder (i.e., preventing or reducing the progression of the disease or alleviating at least one clinical symptom of the disease). In another embodiment, "therapy" or "treatment" refers to improving at least one physical parameter, which may or may not be discernible by a subject, particularly a human subject, that is based on or associated with the disease or disorder being treated. In yet another embodiment, "therapy" or "treatment" refers to modulating or alleviating the disease or disorder physically (e.g., stabilization of discernible and indiscernible symptoms), physiologically (e.g., stabilization of physiological parameters), or both. In yet another embodiment, "therapy" or "treatment" refers to slowing the onset or progression of the disease or disorder. Thus, "therapy" or "treatment" includes any causal treatment of the underlying disease or disorder (i.e., disease modification), as well as any treatment of the signs and symptoms of the disease or disorder (whether or not disease modification occurs), and any alleviation or amelioration of the disease or disorder, or its signs and symptoms. The terms "disease" and "disorder" are used largely interchangeably herein.

[0041] As used herein, the term "effective amount" may refer to a prophylactically effective amount, which is the amount of an active compound or pharmaceutical agent, more specifically a prophylactic agent, that inhibits or delays the onset of disease in a subject as desired by a researcher, veterinarian, physician, or other clinician; or a therapeutically effective amount, which is the amount of an active compound or pharmaceutical agent, more specifically a therapeutic agent, that elicits a biological or medical response in a subject as desired by a researcher, veterinarian, physician, or other clinician, which may include, among other things, alleviation of the symptoms or condition of the disease being treated. Methods for determining therapeutically and prophylactically effective dosages of the pharmaceutical agents taught herein are known in the art. The effective amount may vary depending on the compound, the disease and its severity, and the condition, age, weight, sex, etc., of the subject being treated, particularly a human subject. More specifically, a "therapeutically effective amount" or "therapeutically effective dosage" refers to the amount of the plasma and immunoglobulin-containing composition described herein that, when administered, results in a clinically positive response for the treatment of a subject suffering from an infectious disease. Similarly, a "prophylactically effective amount" or "prophylactically effective dose" refers to the amount of a composition comprising plasma and immunoglobulin described herein that inhibits or delays the onset of clinical symptoms of an infectious disease as sought by a researcher, veterinarian, medical doctor, or other clinician. Those skilled in the art will recognize that terms such as "quantity," "amount," and "level" are synonymous and have clearly defined meanings in the art, and will understand that these terms may refer specifically to an absolute quantification of a composition comprising plasma and immunoglobulin described herein that would be considered an effective amount for a use described herein, or a relative quantification of a composition comprising plasma and immunoglobulin described herein, e.g., the concentration of a composition comprising plasma and immunoglobulin described herein as a function of a subject's body weight. A suitable value or range of values ​​may be obtained from a single subject or from a group of subjects (i.e., at least two subjects).

[0042] It is emphasized that while all of the values ​​and ranges disclosed herein for the compositions comprising plasma and immunoglobulin described herein are suitable for various medical indications or purposes, those skilled in the art will recognize that a particular individual may derive improved benefit from treatment with a composition comprising plasma and immunoglobulin by further optimizing the optimal dosage of said components, taking into account a wide range of parameters, including, but not limited to, the nature and extent of the infection being treated, the subject's sex, the subject's age, weight, other medical indications, nutrition, method of administration, metabolic status, interference or influence of or effectiveness of other active pharmaceutical ingredients, etc. Furthermore, each individual may have a somewhat unique response to the composition comprising plasma and immunoglobulin used.

[0043] As used herein, the term "infection originating in the nasopharyngeal region" shall be interpreted in its broadest sense, i.e., any pathological infection that enters or is transmitted to a subject via the nasopharyngeal region (i.e., nose, ears, mouth, throat, upper respiratory tract), where the causative agent has the ability to be transmitted from a first, affected subject with the infection to a second subject without the infection via respiratory secretions, exhaled breath, mucus, or saliva. The disease caused by the causative agent may be local (i.e., the nasopharyngeal region), directed generally to the lower respiratory system (i.e., including the lungs), systemic (i.e., the nasopharyngeal region is merely a portal for the causative agent to enter the subject's bloodstream, causing systemic disease), or directed to the gastrointestinal system. In the context of the present invention, both symptomatic and asymptomatic infections are contemplated, with symptomatic infections being preferred.

[0044] In certain embodiments, the compositions described herein are used to reduce or completely inhibit the progression of infections originating in the nasopharyngeal region to the lower respiratory tract, to reduce or completely inhibit the progression of infections to the systemic system, and to reduce or completely inhibit the progression of infections to the gastrointestinal system.

[0045] The term "respiratory infection" as used herein is broadly interpreted, i.e., any pathological infection of the respiratory system (i.e., the respiratory tract, including the trachea, bronchi, bronchioles, alveoli, pleura, and pleural cavity) in which the causative agent can be transmitted from a first affected subject with a respiratory infection to a second subject without the respiratory infection via respiratory secretions, such as mucus or saliva. Alternative terms that may be used interchangeably with this term include, but are not limited to, "respiratory infection," "infection of the respiratory tract," and "infection of the respiratory tract," all of which are standard terms in the medical field and known to those skilled in the art. Such respiratory infections may be the result of infections occurring outside the respiratory system. For example, SARS-CoV-2 can invade the intestine and cause secondary respiratory infections. In the context of the present invention, both symptomatic and asymptomatic respiratory infections are contemplated, with symptomatic respiratory infections being preferred. As will be understood, the term "symptomatic" refers to the presence of one or more physical manifestations (i.e., symptoms) of a respiratory infection. Typical symptoms of a (clinically mild) respiratory infection are varied and include, but are not limited to, coughing (phlegm), sneezing, nasal and / or lung congestion, runny nose, sore and / or irritated throat, muscle aches, shortness of breath, wheezing, chest tightness, fever, fatigue, itchy and / or watery eyes, and combinations thereof.

[0046] In certain embodiments, the respiratory infection is an upper respiratory tract infection. In alternative embodiments, the respiratory infection is a lower respiratory tract infection. Respiratory infections are generally routinely classified according to the affected part of the respiratory system and can therefore be upper respiratory tract infections and / or lower respiratory tract infections. Upper respiratory tract infections are primarily localized in the nose, sinuses, pharynx, larynx, or any combination thereof, while lower respiratory tract infections are primarily localized in the bronchi, bronchioles, alveoli, pleura, pleural cavities, or any combination thereof. Many respiratory infections are characterized by the simultaneous presence of pathogens in both the upper and lower respiratory tracts.

[0047] In certain embodiments, the compositions described herein are used to reduce or completely inhibit the progression of upper respiratory tract infections to the lower respiratory tract.

[0048] In certain embodiments, the compositions described herein are used to reduce the initial replication of pathogens in the nasopharynx or upper respiratory tract area, hi even more specific embodiments, the compositions described herein are used to alleviate primary infections in the nasopharynx or upper respiratory tract area.

[0049] In certain embodiments, the compositions described herein are used to prevent, reduce, or completely inhibit the transmission of the causative agent (e.g., a pathogen) of an infectious disease from one subject to another, for example, by preventing infection in an uninfected subject or by inhibiting infection in a recently or newly infected subject by priming the immune system of the subject against the causative agent.

[0050] In certain embodiments, the compositions described herein are used to reduce the amount of a causative agent (e.g., a pathogen such as a virus), preferably the amount of an infectious pathogen, in the subject. The amount of the causative agent may be measured by any means known in the art. For example, the amount of infectious viral particles may be measured using a 50% tissue culture infectious dose (TCID 50 For example, the amount of RNA of the causative agent may be quantified using RT-qPCR.

[0051] In certain embodiments, the causative agent of the infectious disease is a pathogen such as a virus, bacterium, bacterial spore, protozoan, viroid, parasite, yeast, or fungus, preferably a virus. In more specific embodiments, the infectious disease described herein is a viral, bacterial, fungal, or parasitic infection, i.e., an infection whose primary causative agent is a virus, bacterium, fungus, or parasite (i.e., a pathogen), respectively. Obviously, the infectious disease described herein can be caused by multiple causative agents coexisting in a subject. In certain embodiments, the infectious disease is caused by an opportunistic pathogen, and optionally, the infected subject is an immunocompromised subject.

[0052] In certain embodiments, the causative agent of the disease may be a pathogen transmitted via the nasopharyngeal route, regardless of whether the pathogen itself causes a respiratory infection. For example, some pathogens may gain entry through the nasopharynx but result in systemic or gastrointestinal disease.

[0053] In a further embodiment, the infection is a respiratory infection, more particularly a viral respiratory infection. In a further embodiment, the viral respiratory infection is caused by a Riboviridae virus, preferably a Coronaviridae (i.e., a coronavirus), more preferably a Severe Acute Respiratory Syndrome-associated coronavirus, more preferably SARS-CoV, MERS-CoV, or SARS-CoV-2 (which causes COVID-19), and most preferably SARS-CoV-2. In an alternative embodiment, the respiratory infection is a viral respiratory disease caused by a virus selected from the group consisting of influenza virus, respiratory syncytial virus (RSV), parainfluenza virus, metapneumovirus, rhinovirus, coronavirus, adenovirus, and bocavirus.

[0054] Viral respiratory infection pathogens of particular interest in light of the present disclosure are RNA viruses, preferably selected from the group consisting of Bunyaviridae, Coronaviridae, Filoviridae, Flaviviridae, Paramyxoviridae, Picornaviridae, Orthomyxoviridae, and Rhabdoviridae. In a further preferred embodiment, the causative viral pathogen is a coronavirus classified as a coronavirus species selected from the group consisting of alphacoronavirus, betacoronavirus, gammacoronavirus, and deltacoronavirus species. In a further preferred embodiment, the causative viral pathogen is a coronavirus belonging to the genus Coronavirus selected from the group consisting of alphacoronavirus 1, human coronavirus 229E, human coronavirus NL63, miniopterus bat coronavirus 1, miniopterus bat coronavirus HKU8, porcine epidemic diarrhea virus, rhinoceros bat coronavirus HKU2, Scotophilus bat coronavirus 512, betacoronavirus, hedgehog coronavirus 1, human coronavirus HKU1, Middle East respiratory syndrome-associated coronavirus, murine coronavirus, pipistrelle bat coronavirus HKU5, russet bat coronavirus HKU9, severe acute respiratory syndrome-associated coronavirus, Tyronycteris bat coronavirus HKU4, avian coronavirus, beluga whale coronavirus SW1, brown-eared bulbul coronavirus HKU11, and porcine coronavirus HKU15. In a further preferred embodiment, the causative viral pathogen is a betacoronavirus, preferably a betacoronavirus belonging to the subgenus Sarbecovirus.

[0055] In another embodiment, the viral pathogen is an RNA virus, preferably a positive-sense RNA virus, more preferably a positive-sense single-stranded RNA virus.

[0056] The "SARS-CoV-2 virus," commonly referred to as the "COVID-19 virus" and "hCoV-19," is a positive-sense, single-stranded RNA virus that causes respiratory SARS-CoV-2 or COVID-19 disease as described herein. Clearly, when referring to SARS-CoV-2 herein, this term encompasses all documented and undocumented variants of the SARS-CoV-2 virus. According to the common knowledge of those skilled in the art, the term "(genetic) variant," as used herein, refers to a subtype of a pathogen that is genetically distinct from the reference (i.e., "main") genetic strain of said pathogen. Suitable reference SARS-CoV-2 strains are those corresponding to the WIV04 / 2019 sequence (GISAID S lineage, PANGO A lineage, Nextstrain 19B lineage), which is generally referred to as the sequence closest to the original SARS-CoV-2 sequence capable of infecting human subjects (Zhukova et al., "Origin, Evolution, and Global Spread of SARS-CoV-2," CR Biol, 2020). Such variants are also contemplated by the present disclosure. Non-limiting examples of documented SARS-CoV-2 variants include Cluster 5 variants, lineage B.1.1.7 variants, lineage B.1.1.207 variants, lineage B.1.1.317 variants, lineage B.1.1.318 variants, lineage B.1.351 variants, lineage B.1.429 (CAL.20C) variants, lineage B.1.525 variants, lineage B.1.526 variants, lineage B.1.617 variants, lineage B.1.618 variants, lineage P.1 variants, and lineage P.3 variants. Additionally, the term SARS-CoV-2 also includes, but is not limited to, variants characterized by missense mutations selected from the group consisting of D614G, E484K, N501Y, S477G, S477N, P681H, E484Q, L452R, P614R, or any combination thereof.Other non-limiting examples include SARS-CoV-2 isolate Wuhan-Hu-1, alpha variants (also known as the UK variant) (e.g., VOC 202012 / 01, B.1.1.7), gamma variants (also known as the Brazil-Japan variant) (e.g., B.1.1.28 or P1), and beta variants (also known as the South African variant) (e.g., VOC 501Y.V2, B. 1.351), epsilon variants (also known as California variants (e.g., B.1.427 or B.1.429), iota variants (also known as New York variants) (e.g., B.1.526 or B.1.526.1), eta variants (also known as UK / Nigeria variants) (e.g., B.1.525), kappa variants (also known as India variants) (e.g., B.1.617, B.1.617.1, B.1.617.2, or These include variants such as B.1.617.3), Zeta variants (also known as the Brazilian variant) (e.g., P.2), Theta variants (e.g., P3), Lambda variants (e.g., C.37), Mu variants (e.g., B.1.621), Delta variants (e.g., B.1.617.2), or Omicron variants (e.g., B.1.1.529). Skilled researchers are aware of the routine sequencing methods available for detecting genetic variants. Furthermore, both government agencies and research groups provide publicly available repositories containing information on (novel) genetic variants (e.g., https: / / covid.cdc.gov / covid-data-tracker / #variant-proportions and Rakha et al., COVID-19 Variant Database: A Repository of Human SARS-CoV2 Polymorphism Data, BioRxiv, 2020).

[0057] In a preferred embodiment, the SARS-CoV-2 virus is the Wuhan Hu-1, beta, delta, or omicron variant of SARS-CoV-2. In a preferred embodiment, the SARS-CoV-2 virus is the Wuhan SARS-CoV-2 strain BetaCov / Belgium / GHB-03021 / 202 (EPI ISL 407976|2020-02-03) strain.

[0058] In certain embodiments, the causative agent of the infectious disease, such as a respiratory infection, to be treated or prevented in the subject can be a variant or mutant of the causative agent of the respiratory infection to which the donor subject developed a humoral immune response. For example, if the causative agent of the respiratory infection to be treated or prevented in the subject is a beta variant of SARS-CoV-2, the causative agent of the respiratory infection to which the donor subject developed a humoral immune response can be another variant of SARS-CoV-2, such as the Wuhan variant, the Delta variant, or the Omicron variant. Vice versa.

[0059] In an alternative embodiment, the infection, such as a respiratory infection, is a bacterial respiratory infection. In yet another embodiment, the bacterial infection is caused by a bacterium selected from the group consisting of Haemophilus influenzae, Streptococcus pneumoniae, Moraxella catarrhalis, Corynebacterium diphtheriae, Legionella pneumophila, Bordetella pertussis, Coxiella burnetii, Streptococcus pyogenes, Mycobacterium tuberculosis, Mycobacterium avium, Chlamydophila pneumoniae, Chlamydophila psittaci, Chlamydia trachomatis, Klebsiella pneumoniae, Mycoplasma pneumoniae, Pseudomonas aeruginosa, Candida spp., Acinetobacter spp., and Escherichia coli.

[0060] In an alternative further embodiment, the infectious disease, such as a respiratory infection, is a fungal respiratory disease. In a further embodiment, the fungal infection is caused by a fungus selected from the group consisting of Aspergillus, Cryptococcus, Pneumocystis, and endemic mycoses. Exemplary endemic mycoses include, but are not limited to, Blastomyces dermatitidis, Coccidioides immitis, Coccidioides posadasii, Histoplasma capsulatum, Paracoccidioides brasiliensis, Penicillium marneffei, and Sporothrix schenckii.

[0061] In an alternative embodiment, the infection, such as a respiratory infection, is a yeast respiratory infection, such as a yeast lung infection caused by Candida, such as candidiasis.

[0062] In an alternative further embodiment, the infectious disease, such as a respiratory infection, is a parasitic respiratory disease. In a further embodiment, the parasitic infection is caused by a parasite selected from the group consisting of nematodes (e.g., Ascaris lumbricoides, Ancylostoma stercoralis, Strongyloides stercoralis, Mammomonogamus larvae, Dirofilaria immitis, Brugia malayi, Brugia bancrofti, Toxocara canis, Toxocara cati, and Trichinella spiralis), trematodes (e.g., Schistosomiasis, Paragonimiasis), cestodes (e.g., Echinococcus granulosus), and mesofungi (e.g., Rhinosporidium seeburyi).

[0063] Unless otherwise noted, the terms "subject" or "patient" can be used interchangeably and refer to animals, preferably warm-blooded animals, more preferably vertebrates, even more preferably mammals, such as chimpanzees and other apes and monkey species, cows, sheep, pigs, goats, horses, dogs, cats, mice, rats, and guinea pigs, and even more preferably primates, specifically including human patients, non-human mammals, and primates. Preferred subjects are human subjects of all genders and all ages. Both adult subjects, elderly subjects, children, and newborn subjects are intended to be covered by the term "subject." The term "subject" or "patient" includes subjects in need of treatment, more specifically, subjects who would benefit from treatment for a particular condition, particularly an infectious disease. Such subjects may include, but are not limited to, those diagnosed with the condition, those prone to developing the condition, and / or those in need of prevention of the condition. The term "subject" or "patient" includes both singular and plural subjects or patients, unless the context clearly dictates otherwise.

[0064] "Diagnosed," "diagnosing," and "diagnosis" refer to the process of recognizing, determining, or concluding a disease, condition, or (adverse) side effect in a subject based on symptoms and signs and / or the results of various diagnostic procedures (e.g., knowing the presence and / or amount of one or more biomarkers or clinical symptoms characteristic of the diagnosed disease or condition). As taught herein, "diagnosis" of a disease, condition, or (adverse) side effect in a subject may specifically mean that the subject has such disease or condition. A subject may be diagnosed as not having such disease or condition despite exhibiting one or more conventional symptoms or signs associated with such disease or condition. As taught herein, "diagnosis" of a disease or condition in a subject may specifically mean that the subject has an infectious disease. Alternatively, a subject may be diagnosed as not having an infectious disease despite exhibiting one or more conventional symptoms or signs associated with an infectious disease. "Prognosis" in the context of the present invention refers to a prediction of the progression of a respiratory infection in a subject, and the likelihood (e.g., probability, duration, and / or extent) of recovery, and / or the severity or improvement experienced by said infection. The term "good prognosis" generally encompasses the expectation of satisfactory, partial, or complete recovery from a diagnosed pain-inducing disease or condition, optionally within an acceptable period of time. Alternatively, the term may encompass the expectation of further deterioration or no worsening of such condition, preferably within a predetermined period of time. The term "poor prognosis" of a disease or condition typically encompasses the expectation of subpar recovery and / or insufficiently slow recovery, or no recovery at all, or further worsening of the respiratory infection and / or clinical symptoms associated with said disease or condition.

[0065] In the context of the above, "prediction" or "prognosis" generally refers to a statement, declaration, indication, or forecast of a disease or condition in which a subject does not (yet) exhibit any clinical symptoms or limited clinical symptoms or (adverse) side effects. A prediction of a particular clinical disease symptom, condition, or adverse effect in a subject may indicate the probability, likelihood, or risk that the subject will develop that clinical symptom, condition, or (adverse) side effect within a specific period of time, for example, after diagnosis of an infectious disease. This probability, likelihood, or risk may be expressed in any appropriate qualitative or quantitative manner. Non-limiting examples of quantitative expressions include absolute values, ranges, or statistics. Alternatively, the probability, likelihood, or risk may be expressed relative to an appropriate control subject or group of control subjects (i.e., a population of control subjects (e.g., a typical, normal, or healthy subject or subject population)). Therefore, any probability, likelihood, or risk may be advantageously expressed as an increase or decrease, upregulation or downregulation, fold increase or fold decrease, compared to an appropriate control subject or subject population, or compared to a baseline value obtained from a control subject (population) or textbook reference value. When a subject population is used to define a baseline value, the baseline value will obviously be the center size of one or more values ​​(parameters) of the population, such as the mean or median of the values. As those skilled in the art will further understand, monitoring an infectious disease may predict the progression, worsening, remission, or recurrence of the clinical picture or severity of the infectious disease. Furthermore, monitoring may be applied to the course of treatment of a subject. Such monitoring may be involved, for example, in determining whether a patient can be discharged from a controlled clinical or medical setting, whether a change in treatment or therapy is required, or whether (long-term) hospitalization is required.

[0066] In certain preferred embodiments, the subject is a hospitalized subject. In even more preferred embodiments, the subject is a hospitalized subject with a disease considered to have a poor prognosis.

[0067] In certain preferred embodiments, the subject is a non-hospitalized (community treated) patient.

[0068] In certain preferred embodiments, the subject has mild, moderate, or severe symptoms or mild, moderate, or severe clinical symptoms of an infection, such as a respiratory infection, including but not limited to, SARS-CoV-2 infection (COVID-19).

[0069] In certain embodiments, the subject is at high risk of a severe respiratory infection, including but not limited to SARS-CoV-2 infection (COVID-19), and / or may be insufficiently responsive to conventional treatments. Accordingly, in certain embodiments, the subject receiving treatment / prevention of the disease is 60 years of age or older, preferably 65 years of age or older, e.g., 70 years of age or older. In certain embodiments, the subject receiving treatment / prevention of the disease is a subject with severe immune impairment (immunodeficiency). In certain embodiments, the subject receiving treatment / prevention of the disease has a disease or condition selected from the group consisting of hypertension, diabetes, kidney disease, human immunodeficiency virus infection, obesity, Down's syndrome, cardiovascular disease, cancer, and chronic respiratory disease.

[0070] The applicability of the observed technical effects of treatment with the plasma and immunoglobulin-containing compositions described herein is independent of whether the subject receiving the compositions described herein has completed a conventional vaccination procedure against a respiratory infection, such as SARS-CoV-2 (COVID-19).

[0071] Essentially, the compositions of the present invention may be used as a first or rapid response measure to prevent the causative agent from spreading within a population and causing a pandemic, prior to or during the traditional vaccine and / or treatment development process.

[0072] Vaccination (e.g., a conventional vaccine) may not completely prevent the causative agent of the infectious disease from being transmitted from one subject to another. In such cases, it may be advantageous to also administer a composition comprising plasma and immunoglobulins described herein to the vaccinated subject. Thus, in certain embodiments, the subject receiving a composition described herein was vaccinated against the infectious disease (e.g., a conventional vaccine) before receiving a composition comprising convalescent plasma described herein.

[0073] In certain embodiments, the subject receiving the compositions described herein has not been vaccinated against the infectious disease (e.g., a conventional vaccine) prior to receiving the compositions comprising plasma and immunoglobulin described herein. For example, in the early stages of an epidemic, a vaccine (e.g., a conventional vaccine) may not yet be available. The compositions comprising plasma and immunoglobulin described herein allow for treatment of subjects and / or prevention of infection in subjects in the early stages of an epidemic.

[0074] In certain embodiments, administering compositions comprising plasma and immunoglobulins described herein to a subject may be advantageous not only as a first or rapid response measure to prevent the causative agent from spreading through a population and causing a pandemic, but also to induce a humoral immune response in the subject against the causative agent. As a result, compositions comprising plasma and immunoglobulins described herein may be used as early-stage vaccines against the infectious disease.

[0075] In certain embodiments, the infection is a respiratory infection, more particularly, the respiratory infection is SARS-CoV-2, and the subject receiving treatment and / or prevention of the disease does not suffer from chronic olfactory dysfunction (COD).

[0076] In certain embodiments, the donor subject from which the convalescent plasma is collected is from the same species as the subject to whom the composition comprising plasma and immunoglobulin is administered, e.g., if the donor subject is human, the subject to whom the composition comprising plasma and immunoglobulin is administered is also human.

[0077] In certain embodiments, the composition administered to the subject comprises, consists essentially of, or consists of, preferably consists of, plasma and immunoglobulins against the causative agent of the infectious disease, wherein the immunoglobulins are obtained from a donor subject that has developed a humoral immune response against the causative agent of the infectious disease.

[0078] The combination of plasma and immunoglobulin against the causative agent of the infectious disease present in the composition administered to the subject may be convalescent plasma, which essentially comprises a combination of plasma and immunoglobulin, or may be plasma, such as healthy plasma, to which has been added an immunoglobulin, such as purified immunoglobulin against the causative agent of the infectious disease. Preferably, the compositions comprising plasma and immunoglobulin described herein are convalescent plasma.

[0079] Thus, in certain embodiments, when the plasma is combined with purified immunoglobulin, the plasma is not convalescent plasma. In other words, in certain embodiments, when the plasma is combined with purified immunoglobulin, the plasma is not obtained from a donor subject who has developed a humoral immune response against the causative agent of the infectious disease. In certain embodiments, when the plasma is combined with purified immunoglobulin, the plasma is obtained from one or more healthy subjects. A healthy subject may be a subject not affected by a pathological change.

[0080] In certain embodiments, the compositions described herein comprise at least 20% (v / v), at least 25% (v / v), at least 30% (v / v), at least 35% (v / v), at least 40% (v / v), at least 45% (v / v), at least 50% (v / v), at least 55% (v / v), at least 60% (v / v), at least 65% (v / v), at least 70% (v / v), at least 75% (v / v), at least 80% (v / v), at least 85% (v / v), at least 90% (v / v), or at least 95% (v / v), e.g., at least 96% (v / v), at least 97% (v / v), at least 98% (v / v), at least 99% (v / v), or at least 100% (v / v) plasma and immunoglobulins.

[0081] In certain embodiments, when the plasma is combined with purified immunoglobulin, the compositions described herein comprise at least 20% (v / v), at least 25% (v / v), at least 30% (v / v), at least 35% (v / v), at least 40% (v / v), at least 45% (v / v), at least 50% (v / v), at least 55% (v / v), at least 60% (v / v), at least 65% (v / v), at least 70% (v / v), at least 75% (v / v), at least 80% (v / v), at least 85% (v / v), at least 90% (v / v), or at least 95% (v / v), e.g., at least 96% (v / v), at least 97% (v / v), at least 98% (v / v), or at least 99% (v / v) plasma.

[0082] In certain embodiments, when the plasma is combined with purified immunoglobulins, the immunoglobulins (Ig) are purified from convalescent plasma of a donor subject who has developed a humoral immune response to the causative agent of the infectious disease. For example, the immunoglobulins may be purified from convalescent plasma using affinity chromatography, such as affinity chromatography using Protein A (e.g., catalog no. ab270308, Abcam) and Protein G resins (e.g., catalog no. ab270309, Abcam).

[0083] The purified immunoglobulins may be a mixture of immunoglobulins against the causative agent of a respiratory infection and other immunoglobulins present in the blood sample, in certain embodiments, at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, or at least 30% of the total amount of purified immunoglobulin is immunoglobulin against the causative agent of the infection.

[0084] The inventors have found that immunoglobulins present in the purified immunoglobulins but which do not target the causative agent of the infectious disease do not interfere with the functionality of target-specific immunoglobulins that target the causative agent of the infectious disease.

[0085] In certain embodiments, immunoglobulins (Ig) purified from convalescent plasma of a donor subject who has developed a humoral immune response to the causative agent of the infectious disease comprise, consist essentially of, or consist of IgG, IgA, and IgM when the plasma is combined with purified immunoglobulins.

[0086] In certain embodiments, when the plasma is combined with purified immunoglobulin, the compositions described herein contain at least 1 mg / ml, at least 2 mg / ml, at least 3 mg / ml, at least 4 mg / ml, at least 5 mg / ml, at least 6 mg / ml, at least 7 mg / ml, at least 8 mg / ml, at least 9 mg / ml, at least 10 mg / ml, e.g., at least 11 mg / ml, at least 12 mg / ml, or at least 13 mg / ml of Ig obtained from a donor subject that developed a humoral immune response to the causative agent of the infectious disease.

[0087] In certain embodiments, when the plasma is combined with purified immunoglobulin, the compositions described herein comprise at least 100 binding antibody units (BAU) / ml, at least 200 BAU / ml, at least 300 BAU / ml, at least 400 BAU / ml, at least 500 BAU / ml, at least 600 BAU / ml, at least 700 BAU / ml, at least 800 BAU / ml, at least 900 BAU / ml, at least 1000 BAU / ml, at least 1500 BAU / ml, at least 2000 BAU / ml, at least 2500 BAU / ml, at least 3000 BAU / ml, or at least 3500 BAU / ml, preferably at least 450 BAU / ml, more preferably at least 750 BAU / ml, and even more preferably at least 900 BAU / ml of IgG against the causative agent of the infection. In certain embodiments, where the infection is a respiratory infection, for example caused by SARS-CoV-2, the composition comprises at least 100 BAU / ml, at least 200 BAU / ml, at least 300 BAU / ml, at least 400 BAU / ml, at least 500 BAU / ml, at least 600 BAU / ml, at least 700 BAU / ml, at least 800 BAU / ml, at least 900 BAU / ml, at least 1000 BAU / ml, at least 1500 BAU / ml, at least 2000 BAU / ml, at least 2500 BAU / ml, at least 3000 BAU / ml, or at least 3500 BAU / ml, preferably at least 450 BAU / ml, such as at least 600 BAU / ml, more preferably at least 750 BAU / ml, and even more preferably at least 900 BAU / ml of anti-receptor binding domain (RBD) IgG.

[0088] In certain embodiments, when the plasma is combined with purified immunoglobulin, the compositions described herein comprise at least 5 μg / ml, at least 6 μg / ml, at least 7 μg / ml, at least 8 μg / ml, at least 9 μg / ml, at least 10 μg / ml, at least 11 μg / ml, at least 12 μg / ml, at least 13 μg / ml, at least 14 μg / ml, at least 15 μg / ml, at least 20 μg / ml, at least 30 μg / ml, at least 40 μg / ml, at least 50 μg / ml, at least 60 μg / ml, at least 70 μg / ml, at least 80 μg / ml, at least 90 μg / ml, or at least 100 μg / ml, preferably at least 10 μg / ml, and more preferably at least 15 μg / ml, of IgG against the causative agent of the infection. In certain embodiments, where the infection is a respiratory infection, e.g., caused by SARS-CoV-2, the composition comprises at least 5 μg / ml, at least 6 μg / ml, at least 7 μg / ml, at least 8 μg / ml, at least 9 μg / ml, at least 10 μg / ml, at least 11 μg / ml, at least 12 μg / ml, at least 13 μg / ml, at least 14 μg / ml, at least 15 μg / ml, at least 20 μg / ml, at least 30 μg / ml, at least 40 μg / ml, at least 50 μg / ml, at least 60 μg / ml, at least 70 μg / ml, at least 80 μg / ml, at least 90 μg / ml, or at least 100 μg / ml, preferably at least 10 μg / ml, more preferably at least 15 μg / ml, of anti-receptor binding domain (RBD) IgG.

[0089] In certain embodiments, when the plasma is combined with purified immunoglobulin, the compositions described herein comprise at least 100 BAU / ml, at least 200 BAU / ml, at least 300 BAU / ml, at least 400 BAU / ml, at least 500 BAU / ml, at least 600 BAU / ml, at least 700 BAU / ml, at least 800 BAU / ml, at least 900 BAU / ml, at least 1000 BAU / ml, at least 1500 BAU / ml, at least 2000 BAU / ml, at least 2500 BAU / ml, at least 3000 BAU / ml, at least 3500 BAU / ml, at least 4000 BAU / ml, at least 4500 BAU / ml, at least 5000 BAU / ml, or at least 5500 BAU / ml, preferably at least 300 BAU / ml, more preferably at least 400 BAU / ml, of IgA against the causative agent of the infection. In certain embodiments, where the infection is a respiratory infection, for example caused by SARS-CoV-2, the composition comprises at least 100 BAU / ml, at least 200 BAU / ml, at least 300 BAU / ml, at least 400 BAU / ml, at least 500 BAU / ml, at least 600 BAU / ml, at least 700 BAU / ml, at least 800 BAU / ml, at least 900 BAU / ml, at least 1000 BAU / ml, at least 1500 BAU / ml, at least 2000 BAU / ml, at least 2500 BAU / ml, at least 3000 BAU / ml, at least 3500 BAU / ml, at least 4000 BAU / ml, at least 4500 BAU / ml, at least 5000 BAU / ml, or at least 5500 BAU / ml, preferably at least 300 BAU / ml, more preferably at least 400 BAU / ml, of anti-RBD IgA.

[0090] In certain embodiments, when the plasma is combined with purified immunoglobulin, the compositions described herein comprise at least 0.1 μg / ml, at least 0.15 μg / ml, at least 0.2 μg / ml, at least 0.25 μg / ml, at least 0.3 μg / ml, at least 0.35 μg / ml, at least 0.4 μg / ml, at least 0.45 μg / ml, at least 0.5 μg / ml, at least 1 μg / ml, at least 5 μg / ml, at least 7.5 μg / ml, at least 10 μg / ml, preferably at least 0.25 μg / ml, more preferably at least 0.3 μg / ml of IgA against the causative agent of the infection. In certain embodiments, where the infection is a respiratory infection, for example caused by SARS-CoV-2, the composition comprises at least 0.1 μg / ml, at least 0.15 μg / ml, at least 0.2 μg / ml, at least 0.25 μg / ml, at least 0.3 μg / ml, at least 0.35 μg / ml, at least 0.4 μg / ml, at least 0.45 μg / ml, at least 0.5 μg / ml, at least 1 μg / ml, at least 5 μg / ml, at least 7.5 μg / ml, at least 10 μg / ml, preferably at least 0.25 μg / ml, more preferably at least 0.3 μg / ml of anti-receptor binding domain (RBD) IgA.

[0091] In certain embodiments, when the plasma is combined with purified immunoglobulin, the compositions described herein comprise: - at least 5 μg / ml, at least 6 μg / ml, at least 7 μg / ml, at least 8 μg / ml, at least 9 μg / ml, at least 10 μg / ml, at least 11 μg / ml, at least 12 μg / ml, at least 13 μg / ml, at least 14 μg / ml, or at least 15 μg / ml, preferably at least 10 μg / ml, more preferably at least 15 μg / ml, of IgG against the causative agent of said infection; and - at least 0.1 μg / ml, at least 0.15 μg / ml, at least 0.2 μg / ml, at least 0.25 μg / ml, at least 0.3 μg / ml, preferably at least 0.25 μg / ml, more preferably at least 0.3 μg / ml of IgA against the causative agent of said infection Includes:

[0092] In certain embodiments, when the plasma is combined with purified immunoglobulin, the compositions described herein do not contain detectable levels of IgM against a causative agent of a respiratory infection (e.g., anti-RBD IgM when the respiratory infection is caused by SARS-CoV-2). In certain embodiments, when the plasma is combined with purified immunoglobulin, the compositions described herein contain less than 50 BAU / ml, less than 40 BAU / ml, less than 30 BAU / ml, less than 20 BAU / ml, or less than 10 BAU / ml of IgM against a causative agent of the infection, such as anti-RBD IgM when the infection is, for example, a respiratory infection caused by SARS-CoV-2.

[0093] In certain embodiments, when the plasma is combined with purified immunoglobulin, the donor subject of the plasma and the donor subject of the immunoglobulin are different subjects or groups of subjects.

[0094] As used herein, the term "convalescent plasma" refers to plasma obtained from a subject who has developed a humoral immune response to a causative agent of an infectious disease. As a result, this convalescent plasma contains antibodies, e.g., immunoglobulins, against the causative agent at hand. The subject may have acquired humoral immunity by fighting and / or recovering from infection with the causative agent, or by variolation, vaccination, or other means. Convalescent plasma typically contains one or more cytokines (e.g., IL-10, chemokine ligand 2 (CCL2), or tumor necrosis factor (TNF)), one or more downstream effectors of innate immunity, one or more complement activators (e.g., C3 complement), and / or one or more coagulation factors (such as coagulation factors I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, and / or XIII). Preferably, the one or more complement activators and / or coagulation factors are present in convalescent plasma at normal physiological levels (i.e., the physiological concentration of the one or more coagulation factors in a healthy subject).

[0095] Preferably, said convalescent plasma is obtained from a donor subject who has recovered from and / or been vaccinated against said infectious disease.

[0096] Previous infection may be confirmed by any method known in the art, such as, but not limited to, PCR, chest computed tomography, or serological testing. If the infection is caused by a respiratory infection, such as the SARS-CoV2 virus, the donor subject may have received the anti-SARS-CoV2 Pfizer / BioNTech (Co-mirnaty) vaccine and the anti-SARS-CoV2 Moderna (Spikevax) booster vaccine.

[0097] In certain embodiments, the convalescent plasma is obtained from a donor subject who has recovered from the infection and has not been vaccinated against the infection.

[0098] In certain embodiments, the convalescent plasma is obtained from a donor subject who has recovered from the infection and who was vaccinated against the infection prior to contracting the infection.

[0099] In certain embodiments, the convalescent plasma is obtained from a donor subject who has recovered from the infection and whose symptoms have resolved at least 10 days, at least 11 days, at least 12 days, at least 13 days, and preferably at least 14 days prior to collection of convalescent plasma from the donor subject.

[0100] Preferably, the convalescent plasma is allogeneic convalescent plasma, meaning that the donor subject and the subject receiving the pharmaceutical composition comprising the convalescent plasma are from the same species but are genetically distinct.

[0101] Convalescent plasma, as referred to herein, is typically naturally rich in antibodies against the causative agent (e.g., pathogen) of said infection.

[0102] In certain embodiments, the convalescent plasma contains at least 5 mg / ml, at least 6 mg / ml, at least 7 mg / ml, at least 8 mg / ml, at least 9 mg / ml, at least 10 mg / ml, e.g., at least 11 mg / ml, at least 12 mg / ml, at least 13 mg / ml, at least 15 mg / ml, at least 16 mg / ml, at least 17 mg / ml, or at least 18 mg / ml of Ig.

[0103] In certain embodiments, the convalescent plasma comprises at least 100 binding antibody units (BAU) / ml, at least 200 BAU / ml, at least 300 BAU / ml, at least 400 BAU / ml, at least 500 BAU / ml, at least 600 BAU / ml, at least 700 BAU / ml, at least 800 BAU / ml, at least 900 BAU / ml, at least 1000 BAU / ml, at least 1500 BAU / ml, at least 2000 BAU / ml, at least 2500 BAU / ml, at least 3000 BAU / ml, or at least 3500 BAU / ml, preferably at least 450 BAU / ml, more preferably at least 750 BAU / ml, and even more preferably at least 900 BAU / ml of IgG against the causative agent of the infection. In certain embodiments, when the infection is a respiratory infection, for example caused by SARS-CoV-2, the convalescent plasma comprises binding antibody units (BAU) / ml of anti-receptor binding domain (RBD) IgG of at least 100 BAU / ml, at least 200 BAU / ml, at least 300 BAU / ml, at least 400 BAU / ml, at least 500 BAU / ml, at least 600 BAU / ml, at least 700 BAU / ml, at least 800 BAU / ml, at least 900 BAU / ml, at least 1000 BAU / ml, at least 1500 BAU / ml, at least 2000 BAU / ml, at least 2500 BAU / ml, at least 3000 BAU / ml, or at least 3500 BAU / ml, preferably at least 450 BAU / ml, more preferably at least 750 BAU / ml, and even more preferably at least 900 BAU / ml.

[0104] In certain embodiments, the convalescent plasma comprises at least 5 μg / ml, at least 6 μg / ml, at least 7 μg / ml, at least 8 μg / ml, at least 9 μg / ml, at least 10 μg / ml, at least 11 μg / ml, at least 12 μg / ml, at least 13 μg / ml, at least 14 μg / ml, at least 15 μg / ml, at least 20 μg / ml, at least 30 μg / ml, at least 40 μg / ml, at least 50 μg / ml, at least 60 μg / ml, at least 70 μg / ml, at least 80 μg / ml, at least 90 μg / ml, or at least 100 μg / ml, preferably at least 10 μg / ml, more preferably at least 15 μg / ml, of IgG against the causative agent of the infection. In certain embodiments, when the infection is a respiratory infection, e.g., caused by SARS-CoV-2, the convalescent plasma comprises at least 5 μg / ml, at least 6 μg / ml, at least 7 μg / ml, at least 8 μg / ml, at least 9 μg / ml, at least 10 μg / ml, at least 11 μg / ml, at least 12 μg / ml, at least 13 μg / ml, at least 14 μg / ml, at least 15 μg / ml, at least 20 μg / ml, at least 30 μg / ml, at least 40 μg / ml, at least 50 μg / ml, at least 60 μg / ml, at least 70 μg / ml, at least 80 μg / ml, at least 90 μg / ml, or at least 100 μg / ml, preferably at least 10 μg / ml, more preferably at least 15 μg / ml, of anti-receptor binding domain (RBD) IgG.

[0105] In certain embodiments, the convalescent plasma comprises at least 100 BAU / ml, at least 200 BAU / ml, at least 300 BAU / ml, at least 400 BAU / ml, at least 500 BAU / ml, at least 600 BAU / ml, at least 700 BAU / ml, at least 800 BAU / ml, at least 900 BAU / ml, at least 1000 BAU / ml, at least 1500 BAU / ml, at least 2000 BAU / ml, at least 2500 BAU / ml, at least 3000 BAU / ml, at least 3500 BAU / ml, at least 4000 BAU / ml, at least 4500 BAU / ml, at least 5000 BAU / ml, or at least 5500 BAU / ml, preferably at least 300 BAU / ml, more preferably at least 400 BAU / ml, of IgA against the causative agent of the infection. In certain embodiments, when the infection is a respiratory infection, for example caused by SARS-CoV-2, the convalescent plasma comprises at least 100 BAU / ml, at least 200 BAU / ml, at least 300 BAU / ml, at least 400 BAU / ml, at least 500 BAU / ml, at least 600 BAU / ml, at least 700 BAU / ml, at least 800 BAU / ml, at least 900 BAU / ml, at least 1000 BAU / ml, at least 1500 BAU / ml, at least 2000 BAU / ml, at least 2500 BAU / ml, at least 3000 BAU / ml, at least 3500 BAU / ml, at least 4000 BAU / ml, at least 4500 BAU / ml, at least 5000 BAU / ml, or at least 5500 BAU / ml, preferably at least 300 BAU / ml, more preferably at least 400 BAU / ml.

[0106] In certain embodiments, the convalescent plasma comprises at least 0.05 mg / kg, at least 0.1 μg / ml, at least 0.15 μg / ml, at least 0.2 μg / ml, at least 0.25 μg / ml, at least 0.3 μg / ml, at least 0.35 μg / ml, at least 0.4 μg / ml, at least 0.45 μg / ml, at least 0.5 μg / ml, at least 1 μg / ml, at least 5 μg / ml, at least 7.5 μg / ml, at least 10 μg / ml, preferably at least 0.25 μg / ml, more preferably at least 0.3 μg / ml of IgA against the causative agent of the infection. In certain embodiments, when the infection is a respiratory infection, e.g., caused by SARS-CoV-2, the convalescent plasma comprises at least 0.1 μg / ml, at least 0.15 μg / ml, at least 0.2 μg / ml, at least 0.25 μg / ml, at least 0.3 μg / ml, at least 0.35 μg / ml, at least 0.4 μg / ml, at least 0.45 μg / ml, at least 0.5 μg / ml, at least 1 μg / ml, at least 5 μg / ml, at least 7.5 μg / ml, at least 10 μg / ml, preferably at least 0.25 μg / ml, more preferably at least 0.3 μg / ml of anti-receptor binding domain (RBD) IgA. In some embodiments, the amount of IgA against the causative agent of the infection contained in the convalescent plasma is at most 0.1 mg, preferably at most 0.07 mg, per kg of Ig against the causative agent.

[0107] In certain embodiments, the convalescent plasma does not contain detectable levels of IgM against the causative agent of the respiratory infection, e.g., anti-RBD IgM when the respiratory infection is caused by SARS-CoV-2. In certain embodiments, the convalescent plasma contains less than 50 BAU / ml, less than 40 BAU / ml, less than 30 BAU / ml, less than 20 BAU / ml, or less than 10 BAU / ml of IgM against the causative agent of the respiratory infection, e.g., anti-RBD IgM when the respiratory infection is caused by SARS-CoV-2.

[0108] In certain embodiments, where the infection is, for example, a respiratory infection caused by SARS-CoV-2, the convalescent plasma is capable of inhibiting viral replication of a SARS-CoV-2 viral strain, preferably the SARS-CoV-2 Wuhan strain, in vitro, for example in Vero E6 cells.

[0109] In certain embodiments, where the infection is a respiratory infection, e.g., caused by SARS-CoV-2, the convalescent plasma has a half-maximal plasma SARS-CoV-2 neutralizing antibody titer at a plasma dilution of at least 1:500, at least 1:1000, at least 1:2000, at least 1:3000, at least 1:4000, or at least 1:5000. The neutralizing antibody titer may be measured by any method known in the art, such as using a plaque reduction neutralization test (PRNT). The half-maximal neutralizing antibody titer indicates the plasma dilution that resulted in a 50% reduction in SARS-CoV-2 plaques.

[0110] In certain embodiments, the convalescent plasma is collected from the donor subject by plasmapheresis.

[0111] In certain embodiments, the plasma or convalescent plasma is substantially free of cells (i.e., up to 50,000 cells per μL of convalescent plasma, preferably up to 40,000 cells per μL of convalescent plasma).

[0112] In certain embodiments, the plasma or convalescent plasma is a pooled plasma sample, i.e., the plasma or convalescent plasma is obtained from a single sample pool, which includes samples from more than one subject and / or samples from a single subject taken at different time points. In certain embodiments, the plasma or convalescent blood sample is a pooled sample, which includes plasma or convalescent plasma from up to 100, up to 90, up to 80, up to 70, up to 60, up to 50, up to 40, up to 30, up to 20, up to 10, or up to 5 donor subjects. In certain embodiments, the plasma or convalescent blood sample is obtained from a single donor subject.

[0113] In certain embodiments, the plasma or convalescent blood sample is not platelet-rich plasma (PRP). In certain embodiments, the compositions described herein, the plasma or convalescent blood sample comprises less than 100,000 platelets per μl, less than 90,000 platelets per μl, less than 80,000 platelets per μl, less than 70,000 platelets per μl, less than 60,000 platelets per μl, less than 50,000 platelets per μl, less than 40,000 platelets per μl, less than 30,000 platelets per μl, less than 20,000 platelets per μl, less than 10,000 platelets per μl, less than 5,000 platelets per μl, or less than 1,000 platelets per μl.

[0114] In certain embodiments, the compositions described herein comprise at least 1% (v / v), at least 5% (v / v), at least 10% (v / v), at least 15% (v / v), at least 20% (v / v), at least 25% (v / v), at least 30% (v / v), at least 35% (v / v), at least 40% (v / v), at least In certain embodiments, the compositions described herein comprise 45% (v / v), at least 50% (v / v), at least 55% (v / v), at least 60% (v / v), at least 65% (v / v), at least 70% (v / v), at least 75% (v / v), at least 80% (v / v), at least 85% (v / v), at least 90% (v / v), or at least 95% (v / v), e.g., at least 96% (v / v), at least 97% (v / v), at least 98% (v / v), at least 99% (v / v), or 100% (v / v) of convalescent plasma obtained from the donor subject.

[0115] In certain embodiments, the compositions described herein further comprise antibodies, e.g., immunoglobulins, against the causative agent of an infectious disease present in convalescent plasma, or purified immunoglobulins against the causative agent of an infectious disease present in the composition, plus an antibody or antibody fragment against the causative agent of the infectious disease. In certain embodiments, the compositions may further comprise one or more recombinant antibodies or antibody fragments, e.g., recombinant monoclonal antibodies, against the causative agent of the infectious disease. For example, if the infectious disease is the respiratory infection COVID-19, the composition may further comprise an immunoglobulin M (IgM) neutralizing antibody (IgM-14) as described by Zhiqiang Ku et al., "Nasal administration of IgM provides broad protection against SARS-CoV-2 variants." Nature, 2021, 595:718-723.

[0116] As used herein, the term "antibody" is used in its broadest sense and generally refers to any immunological binding agent, such as whole antibodies, including, but not limited to, chimeric, humanized, human, recombinant, transgenic, grafted, and single-chain antibodies, or fusion proteins, conjugates, fragments, or derivatives thereof that contain one or more domains that selectively bind to an antigen of interest. Thus, the term "antibody" includes whole immunoglobulin molecules, monoclonal antibodies, chimeric antibodies, humanized antibodies, human antibodies, or immunologically effective fragments of any of these. Thus, the term specifically encompasses intact monoclonal antibodies, polyclonal antibodies, multivalent (e.g., bivalent, trivalent, or higher) and / or multispecific antibodies (e.g., bispecific or higher specific antibodies) formed from at least two intact antibodies, and antibody fragments, so long as they exhibit the desired biological activity (e.g., the ability to specifically bind to an antigen of interest), as well as multivalent and / or multispecific conjugates of such fragments. The term "antibody" includes not only antibodies produced by methods including immunization, but also any polypeptide (e.g., a recombinantly expressed polypeptide) engineered to contain at least one complementarity-determining region (CDR) capable of specifically binding to an epitope of an antigen of interest. Thus, the term applies to such molecules whether produced in vitro, in cell culture, or in vivo.

[0117] The term "antibody fragment" or "antigen-binding portion" includes a portion or region of a full-length antibody, generally the antigen-binding or variable domain thereof. Examples of antibody fragments include Fab, Fab', F(ab)2, Fv, scFv fragments, V H Domain, V L Domain and V HHThese include single domain (sd) Fvs, diabodies, linear antibodies, single-chain antibody molecules, particularly heavy-chain antibodies, and multivalent and / or multispecific antibodies formed from antibody fragments, such as dibodies, tribodies, and multibodies. The designations Fab, Fab', F(ab')2, Fv, scFv, etc., above, shall have their established meanings in the art. In certain embodiments, the antibody fragment may be a Nanobody®.

[0118] In certain embodiments, the compositions described herein do not comprise one or more recombinant antibodies or antibody fragments, such as recombinant monoclonal antibodies, against the causative agent of said infectious disease.

[0119] In certain embodiments, when the compositions described herein comprise convalescent plasma obtained from a donor subject that has developed a humoral immune response to the causative agent of the infectious disease, the convalescent plasma obtained from the donor subject that has developed a humoral immune response to the causative agent of the infectious disease is not subjected to a purification step to isolate and / or enrich the antibodies or antibody fragments therein.

[0120] In certain embodiments, the plasma and / or convalescent plasma is subjected to pathogen inactivation (also known as pathogen reduction). Pathogen inactivation of blood-derived products is known in the art and can be performed using the Intercept Blood System (CERUS, Concord, CA), Mirasol Pathogen Reduction (Terumo BCT, Denver, CO), or Theraflex UVC (Macopharma, Tourcoing, France).

[0121] In certain embodiments, the plasma and / or convalescent plasma is substantially free of infectious agents (e.g., microorganisms, including bacteria, viruses, and protozoa) that can cause disease in a subject receiving the plasma and / or convalescent plasma (or purified immunoglobulins from the convalescent plasma). In more specific embodiments, the plasma and / or convalescent plasma is substantially free of human immunodeficiency virus (HIV), hepatitis viruses (e.g., hepatitis B or hepatitis C), and Treponema pallidum (i.e., the bacterium that causes syphilis).

[0122] In certain embodiments, the plasma and / or convalescent plasma may be fresh plasma and / or convalescent plasma or may be stored prior to use. In certain embodiments, the plasma and / or convalescent plasma is stored at a temperature between -210°C and 25°C. For example, the plasma and / or convalescent plasma may be stored at a temperature between -196°C and -210°C (e.g., in liquid nitrogen), -80°C (e.g., in a freezer), -20°C (e.g., in a freezer), 4°C (e.g., in a refrigerator), or room temperature. The plasma and / or convalescent plasma may be stored for up to one year or longer (e.g., two years). For example, the plasma and / or convalescent plasma may be stored at a temperature below -30°C for one year or longer. In a further example, the plasma and / or convalescent plasma may be stored at a temperature between -25°C and -30°C for up to six months.

[0123] In certain embodiments, the plasma and / or convalescent plasma may be stored for at least 0.5 hours, at least 1 hour, at least 2 hours, at least 4 hours, at least 6 hours, at least 8 hours, at least 10 hours, at least 12 hours, at least 14 hours, at least 16 hours, at least 18 hours, at least 20 hours, at least 22 hours, at least 24 hours, at least 26 hours, at least 28 hours, or at least 30 hours prior to use.

[0124] In certain embodiments, if the plasma and / or convalescent plasma was frozen prior to use, the convalescent plasma may be thawed after freezing at a temperature of at least 3° C., at least 4° C., at least 5° C., at least 10° C., at least 15° C., at least 20° C., at least 25° C., at least 30° C., at least 35° C., at least 36° C., or at least 37° C. For example, the plasma and / or convalescent plasma may be thawed at 4° C. (e.g., in a refrigerator), at room temperature, or at 37° C. (e.g., in a warm water bath).

[0125] The compositions described herein are administered to the airways or respiratory tract to enhance efficacy against the causative agent of said infection.

[0126] Intranasal or nasopharyngeal administration of the compositions described herein preferably satisfies the nasopharyngeal and / or oropharyngeal mucosa, thereby preventing or delaying infection by the causative agents of said infectious diseases.

[0127] Administration to the respiratory tract or respiratory tract can be accomplished by any recognized or known means, including nasopharyngeal administration, inhalation administration, or intranasal administration. For enhanced efficacy, the neutralizing antibody can be delivered to one or more of the upper and lower respiratory tract, including the nasal cavity, nose, sinuses, throat, pharynx (including the nasopharynx, oropharynx, and laryngopharynx), larynx, trachea, bronchi, and lungs.

[0128] Inhalation refers to delivery to and ingestion of the respiratory tract, particularly in the context of ingesting or administering / being administered a drug or compound, including the compositions described herein. The respiratory tract can include the upper and / or lower respiratory tract. The upper respiratory tract includes the nose, nasal cavities, paranasal sinuses, larynx, and trachea. The lower respiratory tract includes the lungs, airways (bronchi and bronchioles), and air sacs (alveoli). Inhalation can occur through the nose or mouth, or by direct administration to the lower respiratory tract, such as intratracheal administration. Thus, inhalation can include exclusively or primarily nasal, oral, oral inhalation, intratracheal inhalation, and intratracheal instillation. Thus, inhalation provides and is contemplated as a means of administration that results in the compositions described herein being deposited exclusively, specifically, or preferentially in the respiratory tract, including the upper and / or lower respiratory tract.

[0129] The term "intranasal," as used herein, includes, but is not limited to, administering, administering, or occurring in or through the nose or nasal structures. The term "intranasal," as used herein and exemplified as an embodiment in the examples, is not intended to be limited to, or to imply, a limitation to, administration directly or specifically or solely through the nose or nasal passages, particularly to the exclusion of other means of administration by which the compositions described herein are delivered or provided to, deposited in, or distributed into the respiratory system.

[0130] As used herein, the term "naso-oropharyngeal" includes, but is not limited to, administering, administering to, or occurring in or through the nose, oral cavity, or pharynx. The oropharynx is the part of the throat at the back of the mouth behind the oral cavity, including the soft palate, the side and back walls of the throat, the tonsils, and the back (approximately one-third) of the tongue.

[0131] Devices for administration or delivery to the respiratory or airway tract are known and recognized by those skilled in the art and in clinical or medical settings and are applicable to the applications, pharmaceutical compositions, methods, and protocols of use of the present invention. Devices include metered dose spray pumps, hand valve sprayers, intranasal or intranasal oropharyngeal spray devices, sprayers, nebulizers, metered dose inhalers (MDIs), pressurized inhalers, inhalers, intranasal inhalers, nasal spray bottles, unit dose containers, pumps, droppers, squeeze bottles, or two-way devices. The convalescent plasma or pharmaceutical compositions can also be delivered by tube, catheter, syringe, pack tail, pledget, nasal or nasal oropharyngeal tampon, or submucosal injection.

[0132] For example, a container closure system for a nasal spray includes the container and all components responsible for metering, atomizing, and delivering the formulation to a subject. A dose of a composition described herein can be metered by a spray pump in a nasal or oral nasal spray device, or it can be pre-metered during manufacture. A nasal or oral nasal spray unit can be designed for unit dosing or can release multiple metered sprays of a formulation containing a pharmaceutical composition described herein. Nasal sprays are applied to the nasal passages for local and / or systemic effects. Current container closure system designs for inhalation spray drug products include both pre-metered and device-metered presentations, which use mechanical or power-assisted and / or energy from the patient's inhalation to generate the spray plume. Pre-metered presentations contain a previously measured dose or portion of a dose in some type of unit (e.g., a single or multiple blister or other cavity) that is inserted into the device during manufacture or by the patient prior to use. A typical device metered unit has a reservoir containing enough formulation for multiple doses, which is delivered as a metered spray by the device itself when actuated by the patient.

[0133] In certain embodiments, the compositions described herein are in a form suitable for administration to the airway or respiratory tract, for example, a form suitable for reaching the nasopharynx, and optionally the oropharynx.

[0134] Preferably, the compositions described herein are in a form suitable for nasal or nasopharyngeal inhalation and / or oral inhalation, such as in the form of an inhalation solution or suspension.

[0135] In certain embodiments, the compositions described herein are in a form suitable for administration to the nasal or nasopharyngeal mucosa.

[0136] In certain embodiments, the compositions described herein are administered in the form of an aerosol, liquid, or liquid spray.

[0137] In certain embodiments, the composition is a pulmonary aerosol formulation, a nasal or nasopharyngeal oral drop, an oronasal drop, a nasal or nasopharyngeal oral wash, a nasal or nasopharyngeal oral spray, or an oronasal spray. Accordingly, a further aspect provided herein is a nasal, nasopharyngeal, or oronasal spray comprising a composition comprising plasma and immunoglobulin against the causative agent of the infectious disease, wherein the immunoglobulin is obtained from a donor subject that has developed a humoral immune response against the causative agent of the infectious disease. In certain embodiments, the spray is an aerosol spray or a nebulized spray.

[0138] The composition or spray may be formulated into a form suitable for administration using methods known and accepted in the art, medical field, and clinical practice. In certain embodiments, the composition or spray is a pharmaceutical composition. In addition to the plasma and immunoglobulins described herein, the composition or spray may contain one or more excipients (e.g., preservatives (e.g., biostatic agents), antioxidants, antifoaming agents, viscosity modifiers, emulsifiers, suspending agents, buffering agents). Commonly used antifoaming agents are certain alcohols (cetostearyl alcohol), insoluble oils (castor oil), stearates, polydimethylsiloxanes, and other silicone derivatives, ethers, and glycols. Commonly used preservatives include benzoic acid, benzalkonium chloride, thiomersal, chlorbutanol, clobutol, potassium sorbate, and methylparaben. Commonly used buffering agents include glycerin, monopotassium phosphate, dipotassium phosphate, and the like. Commonly used antioxidants include sodium metabisulfite, sodium bisulfite, butylated hydroxytoluene, tocopherol, and the like.

[0139] In certain embodiments, the composition or spray may contain up to 0.001% (w / w) of a preservative. In certain embodiments, the composition or spray is preservative-free.

[0140] The pH of a nasal or nasopharyngeal formulation or spray is important to avoid irritation of the nasal or nasopharyngeal mucosa. In certain embodiments, the pH of the composition or spray is between 4.5 and 7.5, preferably between 5.5 and 6.5.

[0141] Prolonged residence time in the nasal or nasopharyngeal region can generally be achieved by using bioadhesive polymers, microspheres, chitosan, or by increasing the viscosity of the formulation or spray.

[0142] In certain embodiments, the compositions comprising plasma and immunoglobulin described herein are administered to the subject at least once, e.g., once, twice, three times, or four times, preferably once or twice daily. In certain embodiments, the compositions comprising plasma and immunoglobulin described herein are administered to the subject for at least 4 consecutive days, e.g., at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, or at least 14 days.

[0143] Nasal, nasopharyngeal or oronasal formulations are typically administered in small volumes, 25-200 μL, or 50-150 μL, for example 100 μL.

[0144] The compositions described herein can be administered to a subject before or after the subject comes into contact with (i.e., is exposed to) a causative agent (e.g., a pathogen) of an infectious disease. Exposure does not necessarily lead to infection. For example, if a person is exposed to a causative agent for a very short period of time, if the amount of causative agent entering the body is insufficient, or if the body's immune system can quickly fight off the causative agent, exposure is unlikely to lead to infection. Exposure to a causative agent can occur as a result of a subject coming into close contact with an infected subject. Those skilled in the art will understand that the time from exposure to a causative agent to the appearance of symptoms varies depending on the type of infectious disease.

[0145] Thus, in certain embodiments, the compositions described herein are administered to a subject before the subject is exposed to the causative agent of the infectious disease. In certain embodiments, the compositions described herein are administered to a subject at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, or at least 6 months before the subject is exposed to the causative agent of the infectious disease.

[0146] In certain embodiments, the compositions described herein are administered to a subject after the subject has been exposed to the agent causing the infectious disease, e.g., after the subject has come into contact with a subject infected with the agent causing the infectious disease (i.e., to prevent infection and the development of a respiratory illness). Preferably, the compositions described herein are administered to the subject immediately after the subject is notified that they have been exposed to the agent causing the respiratory infection. In certain embodiments, the convalescent plasma is administered to the subject less than 1 hour, less than 2 hours, less than 6 hours, less than 12 hours, less than 18 hours, less than 1 day, less than 2 days, less than 3 days, less than 4 days, less than 5 days, less than 6 days, less than 7 days, less than 2 weeks, less than 3 weeks, or less than 4 weeks after the subject was exposed to the agent causing the respiratory infection. In certain embodiments, the convalescent plasma is administered to the subject at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 2 weeks, at least 3 weeks, or at least 4 weeks after the subject was exposed to the causative agent of the infectious disease.

[0147] In certain embodiments, the compositions described herein are administered to a subject before the subject becomes infected with the infectious disease agent (i.e., to avoid infecting exposed but uninfected subjects). In certain embodiments, the compositions described herein are administered to the subject at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 2 weeks, at least 3 weeks, or at least 4 weeks before the subject becomes infected with the infectious disease agent.

[0148] In certain embodiments, the compositions described herein are administered to a subject after the subject has been infected with the infectious disease agent (i.e., to prevent the onset and / or progression of respiratory disease). In certain embodiments, the compositions described herein are administered to a subject at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 2 weeks, at least 3 weeks, or at least 4 weeks after the subject has been infected with the infectious disease agent.

[0149] Subjects receiving the compositions described herein may be undergoing treatment, prophylaxis, or vaccination against conventional infectious diseases.

[0150] Furthermore, the conventional treatment may be administered simultaneously with or at a different time than the compositions described herein (e.g., before or after administration of the compositions described herein). For example, the subject may already be undergoing conventional treatment before initiating treatment with the compositions described herein.

[0151] Said conventional treatment may be administered intranasally, intranasally oropharyngeal, or orally only, or by other routes of administration such as intravenously or intramuscularly.

[0152] In certain embodiments, the compositions described herein are used as an adjunct to conventional infectious disease treatment, prophylaxis, or vaccination methods.

[0153] In certain embodiments, the compositions described herein are used in the absence of conventional methods for treating, preventing, or vaccinating against infectious diseases.

[0154] As used herein, the term "conventional treatment" can be used interchangeably with "standard treatment" and is intended to refer to any treatment believed by a skilled artisan to have some beneficial effect on an infected patient. Numerous examples of conventional treatments have been described in the art. For example, in embodiments in which the infectious disease is a coronavirus, standard treatment protocols include, but are not limited to, administration of an active pharmaceutical ingredient other than convalescent plasma, supplemental oxygen, mechanical respiratory support, or a combination thereof. In a more preferred embodiment, the plasma and immunoglobulin-containing compositions described herein are used as an adjunct to at least one additional pharmaceutically active agent being administered to a subject for the treatment of the infectious disease. The term "active pharmaceutical ingredient" or "API" as referred to herein is intended to be interpreted in accordance with the World Health Organization's definition of the term: "a substance used in a finished medicinal product (FPP) that exhibits pharmacological activity or otherwise has a direct effect on the diagnosis, cure, mitigation, treatment, or prevention of disease, or that is intended to have a direct effect on the restoration, correction, or modification of physiological function in humans."

[0155] In a further preferred embodiment, the at least one additional pharmaceutical active agent is an antiviral agent and / or an anti-inflammatory agent. As used herein, the terms "antiviral agent," "antiviral drug," or "antiviral therapeutic agent" refer to any pharmaceutical active agent used to treat viral infections. In a preferred embodiment, the antiviral agent is selected from the group consisting of nucleoside / nucleotide reverse transcriptase inhibitors (NRTIs), non-nucleoside reverse transcriptase inhibitors (NNRTIs), integrase inhibitors, entry inhibitors, protease inhibitors (PIs), post-attachment inhibitors, booster drugs (i.e., protease inhibitor stimulators), and any combination thereof. The terms "anti-inflammatory agent," "anti-inflammatory drug," or "anti-inflammatory therapeutic agent" refer to any substance that reduces inflammation or swelling, preferably by inhibiting the activity of one or more cyclooxygenase enzymes, such as COX-1 and COX-2. For example, the anti-inflammatory drug may be a nonsteroidal anti-inflammatory drug (NSAID).

[0156] In an alternative preferred embodiment, the at least one additional pharmaceutically active agent is selected from the group consisting of corticosteroids, dexamethasone, tocilzamab, remdesivir, baricitinib, chloroquine, hydrochloroquine, lopinavir, ritonavir, favipiravir, camostat mesylate, azithromycin, interferons, immunomodulators, recombinant monoclonal antibodies, ivermectin, colchicine, cyclooxygenase (COX-1 and / or COX-2) inhibitors, angiotensin-converting enzyme inhibitors (ACE) inhibitors, or combinations thereof.

[0157] Thus, one skilled in the art will understand that the plasma and immunoglobulin-containing compositions described herein can be administered in combination with any of the pharmaceutical active agents described above, and preferred embodiments of such treatment strategies are described in further detail below.

[0158] Furthermore, the term "conventional treatment" is not intended to refer solely to treatment with pharmacologically active ingredients or direct support of respiratory function, but also encompasses post-acute rehabilitation procedures in a hospital setting, such as physical therapy exercises (Thomas et al., "Physical Therapy Management of COVID-19 in the Acute Hospital Setting: Clinical Practice Recommendations," J Physiother, 2020). In certain embodiments, the compositions comprising plasma and immunoglobulins described herein are used in a post-acute hospital setting in subjects who are also undergoing physical therapy to recover from or assist in recovery from an infection, such as SARS-CoV-2.

[0159] The present invention also encompasses delivery devices suitable for intranasal or nasopharyngeal administration for plasma and immunoglobulin-containing compositions, e.g., convalescent plasma-containing compositions. If nasal or nasopharyngeal (mucosal) administration is desired, the compositions may be in a form dispensed by a spray device, such as a squeeze spray dispenser, a droplet dispenser, a pump dispenser, a nebulizer, or an aerosol dispenser.

[0160] Thus, a further aspect provides a nasopharyngeal, nasal or oronasal spray device comprising a composition comprising plasma and immunoglobulin against the causative agent of said infectious disease, wherein said immunoglobulin is obtained from a donor subject who has developed a humoral immune response against the causative agent of said infectious disease.

[0161] Those skilled in the art will appreciate that the spray device may include a container for containing the composition that has a variable volume size, with the selection of the container's fill volume generally dependent on the intended frequency of use of the compositions described herein.

[0162] Those skilled in the art will understand that the spray device can contain one or more doses of the compositions described herein.For example, the device can be a single-dose, double-dose, or multi-dose delivery system.Preferably, the device is a single-dose delivery system, more preferably a disposable single-dose delivery system.

[0163] The spray device may have a specific spray volume, the selection of which depends on the therapeutic dose.

[0164] The spray device may comprise a spray pump, the selection of which depends on the volume of the composition described herein required to support the dosage.

[0165] In certain embodiments, the nasopharyngeal, nasal, or oronasal spray device comprises a metering spray pump or a mechanical spray pump.

[0166] In certain embodiments, the size of the droplets produced by the spray device is 0.1-300 μm, 10-200 μm, 50-200 μm, or 100-200 μm.

[0167] In certain embodiments, the spray device is an aerosol spray device or nebulizer (e.g., a jet nebulizer, an ultrasonic nebulizer, or a mesh nebulizer). Aerosols are typically pressurized with hydrocarbons.

[0168] For example, the spray device may be an OptiNose liquid delivery nasal spray device or other similar device that delivers medication to both the olfactory and respiratory epithelia of the nasal passages but prevents delivery to the lungs by spraying only when the connection between the nose and lungs is closed. In another example, the spray device may be a pressurized olfactory delivery device, such as the Impel NeuroPharma device, a metered dose nasal spray device, or a unit dose nasal spray device. In yet another example, the spray device may be Aptar Pharma's Unidose (UDS) liquid nasal spray system.

[0169] In certain embodiments, compositions comprising plasma and immunoglobulins described herein can stimulate a humoral immune response and, as a result, can be used as first-line vaccines or vaccine-like compositions. Accordingly, a further aspect provides a method for producing a vaccine or vaccine-like composition comprising combining plasma and a composition comprising immunoglobulins against a pathogen that invades or spreads in the nasopharyngeal region with one or more pharmaceutically acceptable excipients, wherein the immunoglobulins are obtained from a donor subject that has developed a humoral immune response against the pathogen.

[0170] Those skilled in the art will understand that embodiments relating to compositions for use in treating or preventing an infection in a subject are also applicable to the nasopharyngeal, nasal or oral nasal sprays and nasal or oral nasal spray devices described herein, and vice versa.

[0171] While the present invention has been described in conjunction with specific embodiments thereof, many alternatives, modifications, and variations will be apparent to those skilled in the art in light of the foregoing description. Accordingly, it is intended to embrace all such alternatives, modifications, and variations as fall within the spirit and broad scope of the appended claims.

[0172] The aspects and embodiments of the present invention disclosed herein are further supported by the following non-limiting examples. [Example]

[0173] Example 1. Intranasally administered convalescent plasma prevents SARS-CoV-2 infection 1.1 Materials and Methods sample Human samples Plasma samples were collected by plasmapheresis from donors who had recovered from SARS-CoV-2 infection (n = 2) or from a vaccinated convalescent donor (n = 1) and designated COVID-19 convalescent plasma (CCP) and vaccinated COVID-19 convalescent plasma (VCCP), respectively. The CCPs used during the study were designated CCP1 and CCP2. Previous infection was confirmed by PCR, chest computed tomography, or serology. Neutralizing titers in convalescent donors were measured by plaque reduction neutralization test (PRNT) at the Rega Institute for Medical Research (KU Leuven, Belgium) at least 14 days after symptom resolution. VCCPs were collected after high (neutralizing) antibody titers were confirmed by ELISA-based testing at least 14 days after symptom resolution. VCCP donors received the Pfizer / BioNTech (Comirnaty) vaccine and a Moderna (Spikevax) booster vaccine. Plasma samples collected from one donor before the COVID-19 pandemic were used as naive, non-immunized controls and compared with non-immunized human plasma (NIP). ヒト )

[0174] Hamster Sample Pooled non-immune hamster plasma (NIP) ハムスターConvalescent hamster plasma (CCP) was provided by Janvier Laboratories (Le Genest-Saint-Isle, France) and the Rega Institute for Medical Research (University of Leuven, Belgium). ハムスター ) was obtained from the Innovative Research laboratory (Pearl Court Novi, USA).

[0175] Human immunoglobulins in hamster plasma Human immunoglobulins from vaccinated convalescent human plasma samples were purified by affinity chromatography using Protein A (catalog no. ab270308, Abcam) and Protein G resins (catalog no. ab270309, Abcam) according to the manufacturer's instructions. Purified human immunoglobulins were added to phosphate-buffered saline (PBS) pH 7.4 as dialysis buffer (pH 7.4) (referred to as purified hIg) or lyophilized in an Alpha 1-4 LSCbasic from Christ (Osterode am Harz, Germany) and purified using NIP. ハムスター Reconstructed with (NIP ハムスター The purified hIg was collected and analyzed.

[0176] In vitro characterization of human plasma samples SARS-CoV-2 in vitro enzyme-linked immunosorbent assay (ELISA) A 96-well microtiter plate was coated overnight with recombinant SARS-CoV-2 receptor binding domain (RBD) (YP_009724390.1) (1 μg / mL).

[0177] IgG / IgA / IgM isotype ELISA The coated microtiter plates were blocked with assay buffer containing either 1% [wt / vol] bovine serum albumin (BSA) in PBS pH 7.4 for IgG isotype ELISAs or 5% [vol / vol] rabbit serum in PBS for IgA and IgM isotype ELISAs. During the blocking step at room temperature (RT), the plasma samples were diluted in assay buffer: NIP. ヒト The CCP and purified hIg samples were diluted 1 / 200 (IgG ELISA), 15 / 100 (IgA ELISA), or 1 / 10 (IgM ELISA). The CCP and purified hIg samples were diluted 1 / 2000 (IgG ELISA), 1.5 / 100 (IgA ELISA), or 1 / 10 (IgM ELISA). The VCCP was diluted 1 / 2000 (IgG and IgA ELISA) or 1 / 10 (IgM ELISA). Three technical replicates were performed for all samples. Each plate contained a 16-step serial dilution of the CCP sample calibrated to the WHO international standard (NIBSC number: 20 / 136) as a standard (Bazett M et al., "Harnessing the lung's innate anticancer effector function with novel bacterial-derived immunotherapy," Oncoimmunology 7, e1398875 (2018)). After a 1-hour incubation step at room temperature, IgG (Fcγ fragment specific), IgA (α chain specific), or IgM (Fc 5μHorseradish peroxide (HRP)-conjugated rabbit anti-human secondary antibodies (IgG: Catalog No. 309-035-008, IgA: Catalog No. 309-035-011, IgM: Catalog No. 309-035-095, Jackson ImmunoResearch) specifically targeting either the IgG or IgM fragments were incubated at room temperature for 1 hour. For IgG isotype ELISAs, a 1:15,000 dilution (5.33 ng / well) was incubated in assay buffer. For IgA and IgM isotype ELISAs, a 1:10,000 dilution (8 ng / well) or a 1:8,000 dilution (10 ng / well) was incubated in assay buffer, respectively. Color development was performed with 3,3',5,5'-tetramethylbenzidine (TMB) substrate solution (catalog no. T444, Sigma-Aldrich, St. Louis, MO, USA) for 6 minutes (IgG), 15 minutes (IgA), or 7 minutes (IgM). Optical density was measured at 450 nm using a spectrophotometer (Plate Reader Infinite F200 PRO, TECAN, Männedorf, Switzerland). For all isotype ELISAs, the intra- and interassay coefficients of variation (CV) were less than 10% and 15%, respectively, for five replicates.

[0178] Concentrated IgG / IgA ELISA Blocking, sample dilution, and secondary antibody dilution were performed as previously described for the IgG / IgA isotype ELISA. Antibody concentrations were measured using anti-Spike-RBD human IgG1 (catalog no. srbd-mab1, Invivogen) spiked into a 1 / 200 human plasma pool (n = 15) or 15% NIP. ヒト Serial dilutions of anti-Spike-RBD human IgA1 (catalog no. srbd-mab6, Invivogen) in PBS were included as a standard. All samples were run in triplicate with technical replicates unless otherwise specified. Color development was performed in TMB for 6 minutes (IgG) or 5 minutes (IgA). For both isotype-enriched ELISAs (n=4), intra- and interassay CVs were less than 15% and 20%, respectively.

[0179] Inhibition ELISA The AcroBiosystems inhibition ELISA (catalog no. EP-105, AcroBiosystems) was performed according to the manufacturer's instructions, with the exception of sample preparation (Almeida JR et al., "A Novel Approach to Overcome Antibiotic Resistance Utilizing Snake Venom Phospholipase A2," Drug Dev Res 80, 68-85 (2019)). Briefly, RBD was coated overnight at 4°C. Plate blocking was performed with 2% [wt / vol] BSA in PBS. Plasma samples were serially diluted 2-fold in PBS containing 0.5% [wt / vol] BSA. The same CCP sample as used in the IgG / IgA / IgM isotype ELISA was included on each plate as a standard to allow conversion to international WHO standard units. Biotinylated angiotensin-converting enzyme 2 (ACE2 receptor) and the above plasma dilutions were mixed in a 1 / 2 volume per well. After incubation, streptavidin-HRP was added. Color development (10 min) and optical density readings were performed as previously described for the IgG / IgA / IgM isotype ELISA.

[0180] Data analysis Data were processed using GraphPad Prism version 9 (GraphPad Software Inc., San Diego, CA, USA). For isotype, concentration, and inhibition ELISAs, background signals (assay buffer or human plasma pool) were subtracted from the raw data. Neutralizing antibody levels measured by isotype and inhibition ELISAs were calibrated against an international WHO standard (NIBSC number: 20 / 136) and expressed as binding antibody units per mL (BAU / mL) and international units per mL (IU / mL), respectively. IgG and IgA concentrations were expressed in μg / mL, the units of a standard (anti-Spike-RBD human IgG1 or IgA1) of known concentration.

[0181] Plaque Reduction Neutralization Test (PRNT) PRNT was performed at the Rega Institute for Medical Research (KU Leuven) as previously described (Betrains A et al., "Convalescent plasma treatment of persistent severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in lymphoma patients with impaired humoral immunity and lack of neutralizing antibodies," Br J Haematol 192, 1100-1105 (2021); Boudewijns R et al., "STAT2 signaling inhibits viral spread but causes severe pneumonia in SARS-CoV-2-infected hamsters," Nature communications 11, 1-10 (2020); Wouters E et al., "A novel competitive ELISA for rapid quantification of SARS-CoV-2 neutralizing antibodies in convalescent plasma," Transfusion 61, 2981-2990 (2021)). The variants of concern tested by PRNT include the Wuhan SARS-CoV-2 strain (BetaCov / Belgium / GHB-03021 / 202, EPI ISL 407976|2020-02-03, passage 5) (Spiteri G et al., "First cases of coronavirus disease 2019 (COVID-19) in the WHO European Region, January 24 to February 21, 2020," Eurosurveillance 25, 2000178 (2020)), and Delta B.1.351 (hCoV-19 / Belgium / rega-1920 / 2021; EPI_ISL_896474, 2021-01-11, passage 2) (Park YJ et al., "Antibody-mediated broad sarbecovirus neutralization by ACE2 molecular mimicry," Science 375, 449-454 (2022)), and Omicron B.1.617.2 (passage 2). Briefly, for all SARS-CoV-2 variants (Wuhan, Delta, Omicron), VCCP, CCP, and NIP were identified. ヒトTen-fold serial dilutions of the SARS-CoV-2 mutants were prepared in duplicate. Dose-dependent neutralization of these test samples was assessed by mixing the plasma dilutions with 100 plaque-forming units (PFU) of the SARS-CoV-2 mutants in DMEM supplemented with 2% FBS. The mixture was incubated at 37°C for 1 hour and then added to a VeroE6 cell monolayer (African green monkey kidney, ATCC CRL-1586) in a 12-well plate. After 1 hour of incubation at 37°C, the inoculated mixture was replaced with 0.8% (w / v) methylcellulose in DMEM supplemented with 2% FBS. After 3 days of incubation at 37°C, the upper layer was removed and the cells were fixed with 3.7% PFA. The cells were stained with 0.5% crystal violet. The half-maximal neutralization titer (PRNT50) was defined as the plasma dilution resulting in a 50% reduction in plaques.

[0182] animal Wild-type Syrian golden hamsters (Mesocricetus auratus) were purchased from Janvier Laboratories (Le Genest-Saint-Isle, France). Six- to eight-week-old female hamsters were used throughout the study. They were specific pathogen-free (SPF). The housing environment and experimental procedures were approved by the Animal Experimentation Ethics Committee of the Catholic University of Leuven (license P065-2020). Food and water were provided ad libitum, along with cage enrichment (wooden blocks). The animals were allowed to acclimate for 4 days before the start of the study.

[0183] SARS-CoV-2 infection model in hamsters The direct contact hamster transmission model has been previously described (Boudewijns R et al., "STAT2 signaling limits viral spread but causes severe pneumonia in SARS-CoV-2-infected hamsters," Nature communications 11, 1-10 (2020); Kaptein SJ et al., "High-dose favipiravir, but not hydroxychloroquine, exhibits potent antiviral activity in SARS-CoV-2-infected hamsters," Proceedings of the National Academy of Sciences 117, 26955-26965 (2020); Sanchez-Felipe L et al., "A single-dose live-attenuated SARS-CoV-2 vaccine candidate in the YF17D vector," Nature 590, 320-325 (2021)). Briefly, index hamsters were inoculated with 2 × 10 6 TCID 50 Hamsters were infected intranasally with 50 μL (25 μL per nostril) containing SARS-CoV-2 (day 0). Virus titers were measured by endpoint dilution on Vero E6 cells using the Reed and Muench method (REED LJ and MUENCH H., "A Simple Method for Estimating the 50th Percent Endpoint," American Journal of Epidemiology 27, 493-497 (1938)). Naive, untreated hamsters (sentinels) were housed with infected hamsters (indexes) in ventilated isolator cages (IsoCage N Biocontainment System, Tecniplast).

[0184] Treatment Schedule Sentinels were treated daily under isoflurane anesthesia for 5 consecutive days, beginning 1 day before exposure to the index hamster, with human or hamster plasma samples, depending on the blinded study design. Treatment groups were administered intranasally (25 μL per nostril). First, sentinels were treated with human plasma samples: CCP (n = 18 in two independent experiments), VCCP (n = 6), purified hIg (n = 6), or NIP. ヒト (n=6). Sentinel treatment with hamster plasma samples was performed with CCP. ハムスター (n=6), NIP ハムスター (n=6), and NIP ハムスター As a negative control, sentinels were treated with PBS pH 7.4 buffer (n=14 in two independent experiments).

[0185] SARS-CoV-2 infection model The SARS-CoV-2 hamster infection model has been previously described (Boudewijns R et al., "STAT2 signaling inhibits viral spread but causes severe pneumonia in SARS-CoV-2-infected hamsters," Nature communications 11, 1-10 (2020); Sanchez-Felipe L et al., "A single-dose live-attenuated YF17D-vectored SARS-CoV-2 vaccine candidate," Nature 590, 320-325 (2021)). Briefly, wild-type Syrian golden hamsters were housed singly in ventilated isolator cages (IsoCage N Biocontainment System, Tecniplast). The experiment was performed at the Rega Institute for Medical Research (KU Leuven).

[0186] Treatment schedule and viral infection The study was conducted in a blinded fashion over a 5-day period at the Rega Institute for Medical Research (KU Leuven). Eighteen hamsters were equally divided into three treatment groups: CCP, buffer control (PBS), and monoclonal anti-RBD neutralizing antibody (catalog number 40592-MM57, Sino Biological).

[0187] 10 for all animals 3 TCID 50 Hamsters were inoculated with SARS-CoV-2 and treated as described in the "SARS-CoV-2 infection model in hamsters" section. This treatment was performed 1 day and 1 hour before virus inoculation. Hamsters were kept without further treatment for 4 days. Hamsters were sacrificed on the 4th day.

[0188] Sample collection Sentinel hamsters were sacrificed on day 4 postchallenge, and index hamsters were sacrificed on day 4 postchallenge. Hamsters were euthanized with an intraperitoneal (ip) injection of 500 μL of Dolesal (200 mg / ml pentobarbital sodium, Vetoquinol SA). Blood samples were collected intracardially using K2 EDTA (BD Vacutainer) blood collection tubes. To obtain plasma, the blood samples were centrifuged at 4000 g for 5 minutes, and the supernatant was collected. For infected hamsters, the plasma was UV-inactivated under a UV lamp (250 nm) under laminar flow for 1 hour. Bronchoalveolar lavage (BAL) fluid was collected by aspirating 500 μL of PBS injected intratracheally. Finally, the BAL samples were centrifuged at 2000 g for 5 minutes, and the supernatant was collected. The entire left lung lobe and nasal epithelium were collected for histopathological and immunofluorescence examination. A portion of the right lung lobe was collected for quantification of viral RNA and infectious virus.

[0189] RNA extraction and quantitative reverse transcription PCR (RT-qPCR) SARS-CoV-2 RT-qPCR has been previously described (Kaptein SJ et al., "High-Dose Favipiravir, but Not Hydroxychloroquine, Exhibits Potent Antiviral Activity in SARS-CoV-2-Infected Hamsters," Proceedings of the National Academy of Sciences 117, 26955-26965 (2020)). Briefly, lung tissue was collected after sacrifice and homogenized in 350 μL of TRK lysis buffer (EZNA® Total RNA Kit, Omega Bio-tek) using a bead breaker (Precellys). Cell debris was pelleted by centrifugation at 10,000 g for 5 minutes, and RNA was extracted according to the manufacturer's instructions. Finally, 50 μL of eluate was collected, of which 4 μL was used as template for RT-qPCR reactions. RT-qPCR was performed using a LightCycler96 platform (Roche). The SARS-CoV-2 nucleocapsid was targeted with the N2 primer and probe using the iTaq Universal Probes One-Step RT-qPCR Kit (BioRad) (Boudewijns R et al., "STAT2 signaling limits viral spread but causes severe pneumonia in SARS-CoV-2-infected hamsters," Nature Communications 11, 1-10 (2020)). A standard of known concentration of SARS-CoV-2 cDNA (from IDT) was used to express the amount of viral genome copies per milligram of tissue. A standard curve of Ct versus genome copies per milliliter was created using 10-fold dilutions of SARS-CoV-2 cDNA. The total number of genome copies was then calculated based on the elution volume of the RNA extract. The number of genome copies was normalized to the weight of the lung tissue from which RNA was extracted.

[0190] Quantification of SARS-CoV-2 infectious particles in lung tissue (TCID 50 ) Endpoint virus titration has been previously described (Kaptein SJ et al., "High-dose favipiravir, but not hydroxychloroquine, exhibits potent antiviral activity in SARS-CoV-2-infected hamsters," Proceedings of the National Academy of Sciences 117, 26955-26965 (2020)). Briefly, lung tissue was homogenized in 350 μL of minimal essential medium (MEM) using a bead breaker (Precellys) and centrifuged at 10,000 g for 5 minutes to pellet cellular debris. Infectious particles were quantified by endpoint titration on confluent Vero E6 cells in 96-well plates. Viral titers were calculated using the Lindenbach calculator by the Reed and Muench method (REED LJ and MUENCH H., "A Simple Method for Estimating the 50 Percent End Point," American Journal of Epidemiology 27, 493-497 (1938)). Viral titers were expressed as 50% tissue culture infectious doses (TCID) per mg of tissue. 50 ) was expressed as

[0191] Histological evaluation Lung histological examination has been previously described (Kaptein SJ et al., "High-Dose Favipiravir, but Not Hydroxychloroquine, Shows Potent Antiviral Activity in SARS-CoV-2-Infected Hamsters," Proceedings of the National Academy of Sciences 117, 26955-26965 (2020)). For histological examination, lungs were fixed overnight in 4% formaldehyde and embedded in paraffin. Longitudinal tissue sections (5 μm) were analyzed after H&E staining, and lung injury was blindly scored by an expert pathologist. Scoring parameters, with a cumulative score of 1–3, included congestion, intraalveolar hemorrhage, apoptotic bodies in the bronchial epithelium, necrotizing bronchiolitis, perivascular edema, bronchopneumonia, perivascular inflammation, peribronchial inflammation, and vasculitis.

[0192] statistics GraphPad Prism version 9 (GraphPad Software Inc., San Diego, CA, USA). Statistical significance between treatment groups was determined by nonparametric Kruskal-Wallis with Dunn's post-hoc test. P values ​​<0.05 were considered significant.

[0193] 1.2 Results Intranasal administration of human (convalescent) plasma in a SARS-CoV-2 hamster infection model In vitro characterization of human plasma samples VCCPs contained 3928 ± 405.9 BAU / mL of anti-RBD IgG and 5949 ± 592.2 BAU / mL of anti-RBD IgA (Figure 1a, b). As expected, CCPs contained significantly less anti-RBD IgG and anti-RBD IgA, at 926.2 ± 101.6 BAU / mL and 450.2 ± 19.56 BAU / mL, respectively. Anti-RBD IgM levels were near or below the detection limit of the assay (Figure 1c). Anti-RBD IgG concentrations were 104.6 ± 10.19 μg / mL in VCCPs and 20.06 ± 0.858 μg / mL in CCPs (Figure 1e). The concentrations of anti-RBD IgA were 12.15 ± 0.25 μg / mL in VCCPs and 0.447 ± 0.045 μg / mL in CCPs (Fig. 1f). All convalescent samples inhibited viral replication of the Wuhan strain in Vero E6 cells in vitro (Fig. 1h). Consistent with the serological data above, the PRNT of this assay was 50 The highest level was observed in VCCP (1:5,000), 10 times higher than in CCP (1:1,000), and in human non-immune plasma (NIP). ヒト These data were consistent with the inhibition of binding of the recombinant RBD to ACE2 in ELISA (Fig. 1g).

[0194] To reduce non-humoral plasma components, the immunoglobulin fraction (hIg) of VCCP was purified using protein A and G affinity chromatography. Purity was qualitatively assessed by SDS-PAGE using Coomassie staining (data not shown). The purified protein content was quantified by BCA assay, and the total protein concentration was 14.8 mg / mL. The anti-RBD IgG concentration was 94.89 ± 13.51 μg / mL (Figure 1e), indicating that 0.6% of the total protein was anti-RBD IgG. Aliquots of the pre-elution, flow-through, and post-elution samples were tested by ELISA to determine the yield of neutralizing antibodies (data not shown). ELISA demonstrated that all fractions still inhibited RBD binding to ACE2, with no difference in the inhibitory levels between VCCP and hIg compared with the significant reduction in the flow-through sample. The levels of anti-RBD IgG were similar to those in VCCP, but the levels of IgA were significantly reduced to 459.1 ± 11.76 BAU / mL (Fig. 1a, b) or 0.236 μg / mL ± 0.019 μg / mL (Fig. 1f).

[0195] Convalescent plasma samples were also screened in vitro for neutralization of Delta and Omicron virus variants infecting Vero E6 cells (Figure 3). As expected, neutralization of the virus was reduced compared to the Wuhan strain. PRNT of Delta variants 50 The ratios were 1:2,000 for VCCP and 1:250 for CCP. 50 The ratio was 1:1,000 for VCCP and 1:100 for CCP. ヒト The samples were not inhibitory. The CCP donor was most likely infected with the Wuhan strain, but the origin of the VCCP cannot be traced. Based on the timing of the donation and the course of the pandemic, it is almost certain that the VCCP was not to Omicron.

[0196] In vivo testing using SARS-CoV-2 infection models We tested the preventive efficacy of intranasally administered CCP in a hamster model of SARS-CoV-2 infection. Sentinel hamsters were treated once daily for 5 consecutive days with either CCP, VCCP, purified human immunoglobulin (hIg), nonimmune plasma (NIP), or buffer. This treatment regimen for sentinel hamsters began on day 0, and index hamsters were separately infected with the SARS-CoV-2 Wuhan strain. One day later, the sentinel and index hamsters were paired and housed together in a cage (Figure 1i). The index and sentinel hamsters were euthanized on days 4 and 5, respectively. Lung tissue was prepared for analysis as previously described (Kaptein SJ et al., "High-dose favipiravir, but not hydroxychloroquine, exhibits potent antiviral activity in SARS-CoV-2-infected hamsters," Proceedings of the National Academy of Sciences 117, 26955-26965 (2020)). No significant weight loss or signs of toxicity were observed throughout the treatment period of the in vivo study (data not shown).

[0197] Treatment with VCCP, CCP, and purified hIg resulted in average viral RNA titers of 2.1, 2.7, and 2.5% in the lungs of sentinel hamsters compared with buffer control. 10 The NIP activity was significantly reduced by log / mg tissue (Fig. 1j). ヒト Treatment with 1.1% CI 1.1% compared to buffer control 10 Although there was a non-significant decrease in log / mg tissue (P=0.1963), VCCP and CCP had a 3.1 and 2.5 log / mg tissue loss, respectively, compared to the buffer control. 10 log TCID 50 A decrease in infectious virus / mg tissue was observed, which was related to the dose of anti-RBD immunoglobulin found in both convalescent plasma samples (Fig. 1k). The infectious virus in the lungs of hamsters treated with purified hIg was 2.4 times higher than that in the buffer control. 10 There was a log / mg tissue reduction, but this was less than the VCCP from which it was derived. ヒトThe lungs of hamsters treated with were not as protected as those of buffer-treated controls (Fig. 1d, l).

[0198] The data were scattered around the median, suggesting variable efficacy of in-treatment with convalescent plasma. However, when stratified by lung infectious viral load, virus was undetectable in 2 of 6 hamsters (33%) in the VCCP group and 7 of 18 hamsters (39%) in the CCP group. Significant reductions in infectious viral load were observed in 4 of 6 hamsters (67%) in the VCCP group and 7 of 18 hamsters (39%) in the CCP group. The remaining 0 of 6 hamsters in the VCCP-treated group and 4 of 18 hamsters (22%) in the CCP-treated group were considered "fully infected," i.e., had the same infectious viral load as the index hamster at the time of euthanasia.

[0199] NIP ヒト The histopathological score of lung tissue from hamsters treated with NIP was 6.8 ± 2.6 (median ± IQR), the highest of all groups. The lung score of the fully infected buffer control group was 4.0 ± 1.6, and the NIP ヒト The lung scores of the CCP group (4.0 ± 2.3) and the VCCP group (3.0 ± 2.9) were not significantly lower than those of the buffer control group (Figure 1l, data not shown). Surprisingly, the lung scores of the CCP group (4.0 ± 2.3) and the VCCP group (3.0 ± 2.9) were not significantly lower than those of the buffer control group. However, the lung scores of the human convalescent plasma (VCCP, CCP, or NIP) were significantly lower than those of the buffer control group (Figure 1l, data not shown). ヒト The type of lung tissue damage observed in hamsters treated with HIV-1 was distinct from the typical infectious inflammation caused by SARS-CoV-2 infection (data not shown). Instead, alveolar epithelial cell hyperplasia and intraalveolar eosinophils were suggestive of allergic toxicity (data not shown). This characteristic histopathological picture was not observed in any of the infected index hamsters, nor in any of the sentinel hamsters treated with buffer (data not shown) or purified hIg (data not shown).

[0200] In vivo studies using SARS-CoV-2 infection models We hypothesized that continuous daily infusion of human plasma induces specific pulmonary clinical features. Therefore, we tested the preventive effect of infusion of CCP in a targeted, rather than transmission, SARS-CoV-2 infection model. In this model, hamsters were administered CCP, NIP, and IV. ヒト Mice were treated twice with either the RBD inhibitor or a control anti-RBD mAb, followed by manual inoculation with virus (Fig. 4a). CCP treatment was performed 24 hours and 1 hour before infection. Euthanasia was performed on day 4 post-infection. The CCP in this study (CCP2) contained anti-RBD IgG at 630.4 BAU / mL ± 125.7 BAU / mL and anti-RBD IgA at 258.7 ± 10.04 BAU / mL, which were 1.5- and 1.7-fold lower than those in CCP1 (Fig. 1a, b and Fig. 4b, c). Anti-RBD IgM antibodies were not detected (Fig. 1c and Fig. 4d). The concentrations of anti-RBD IgG (15.72 ± 0.70 μg / mL) and IgA (0.306 ± 0.028 μg / mL) in CCP2 were similar to those in CCP1 (Fig. 4f, g). ELISA showed similar neutralization levels (i.e., 471.9 IU / mL ± 6.14 IU / mL) for both CCP1 and CCP2 (Figures 1g and 4h). A control mAb was used at a concentration of 30 nM, which inhibited Wuhan virus infection of Vero E6 cells in vitro by 92% (Figure 4e). This concentration was 3.5-fold lower than the 104 nM IgG concentration seen with CCP2 (Figure 4g).

[0201] No significant weight loss or obvious changes in vital signs were observed in any of the cohorts during the experiment (Fig. 4i). Viral RNA levels in lung tissue did not differ significantly between cohorts (Fig. 4j). However, infectious virus in the lungs of animals pretreated with CCP2 was significantly higher than that of NIP. ヒト The expression of NIP was 10-fold lower compared to that of NIP (P = 0.042) (Fig. 4k). ヒトThe infectious virus levels were similar to those of the control group, and therefore no protection was observed. Lung scores did not differ between the groups. There was also no evidence of alveolar epithelial cell hyperplasia or intraalveolar eosinophilia. This suggests that reducing the number of consecutive interventions avoided allergic toxicity (Figure 4l, data not shown).

[0202] In vivo studies using naive hamsters treated in plasma The above data suggest toxicity after repeated administration of human plasma. To assess cross-species efficacy, lung histopathological scoring was performed in a safety study without viral infection after 5 consecutive days of administration using human or hamster (convalescent) plasma or serum (Fig. 5a). Controls were hIg in buffer, hamster plasma, and hIg in buffer. Convalescent or non-immunized human plasma and serum increased cumulative lung scores compared with the buffer control (P = 0.020, n = 3 + 3 + 3) (Fig. 5b, data not shown). NIP ヒト In two-thirds of the hamsters treated with NIP, hyperplastic alveolar epithelial cells were present in only 10% of the lungs (Fig. 5c, data not shown). Purified hIg in hamster plasma resulted in a slight, but not significant, increase in median values ​​of 2.0 ± 1.8 compared with buffer controls (P = 0.108) (data not shown). Most importantly, however, no lung disease was observed in animals treated with hamster-derived plasma or serum, suggesting that interspecies effects play an important role (data not shown). Qualitative analysis of lung sections revealed that NIP ヒト Perivascular edema was observed in 2 / 3 of the hamster plasma hIgs, 1 / 6 of the hamster plasma hIgs, and 1 / 3 of the human serum hIgs, but was absent in the intraspecific treatment cohort and buffer control (Fig. 5, data not shown). Intravesicular hemorrhage was rare but was present in 1 / 6 of the hamster plasma hIgs and 2 / 6 of the buffer hIgs, but was absent in the intraspecific treatment cohort or buffer control (Fig. 5c, data not shown).

[0203] These experiments were conducted to examine the cross-species effects on lung histopathological scoring in a safety study without viral infection. Figure 5 does not demonstrate that plasma and serum can be used interchangeably in the prevention or treatment of disease caused by causative agents that invade or spread to the nasopharyngeal region of subjects.

[0204] Intranasal administration of hamster (convalescent) plasma in a SARS-CoV-2 hamster infection model Because the most likely cause of alveolar epithelial cell hyperplasia and eosinophil infiltration was the repeated administration of human plasma, the next experiment focused again on infection of hamsters administered hamster CCP or hIg (non-immune) in hamster plasma. Non-immune hamster plasma and buffer were used as controls. A similar protocol to that shown in Figure 1 was used.

[0205] In vitro characterization of hamster plasma samples Hamster CCP and hamster plasma hIg inhibited recombinant RBD binding to ACE2 to a similar extent at 4626 IU / mL ± 42.67 IU / mL and 6804 IU / mL ± 111.0 IU / mL, respectively. Non-immune hamster plasma inhibited binding to ACE2 to a similar extent at 50.01 IU / mL ± 8.21 IU / mL, which was less potent than non-immune human plasma (Fig. 2a).

[0206] In vivo studies using SARS-CoV-2 infection models No significant weight loss or obvious changes in vital signs were observed in any of the cohorts during the experiment (Fig. 2b). Viral RNA titers in the lungs of sentinel hamsters treated with hamster CCP or hIg in hamster plasma were 3.1 and 4.5 times higher than those of the buffer control. 10 log TCID 50 / mg tissue was reduced (Fig. 2c). Non-immunized hamster plasma was less effective than the buffer control (P = 0.2056). Infectious virus in the lungs was reduced by 2.7% in hamster CCP compared with the buffer control. 10 log TCID 50 / mg lung, hIg 2.3 in hamster plasma 10 log TCID 50 The lung counts were significantly reduced (Figure 2d). Nonimmunized hamster plasma was less effective than the buffer control (P > 0.999). Categorically, 1 / 6 (17%) of hamsters treated with hamster CCP and 2 / 6 (33.3%) of hamsters treated with hIg in hamster plasma had no detectable virus in their lungs. 3 / 6 (50%) of hamsters treated with hamster CCP and 4 / 6 (66.6%) of hamsters treated with hIg in hamster plasma had significant reductions in virus compared with their corresponding index animals. The remaining 2 / 6 (33%) of the hamster CCP cohort showed infectious virus levels similar to their corresponding index hamsters, and the hIg cohort did not show any infectious virus levels similar to their corresponding index hamsters.

[0207] hIg and NIP in hamster plasma ハムスター The histopathological scores of lung tissue from hamsters treated with CCP were 3.0 ± 3.0 and 3.8 ± 0.8 (median ± IQR), which were not significantly different from the buffer control (4.0 ± 1.6, P = > 0.999) (Fig. 2e, data not shown). ハムスター The lung score in hamsters treated with CCP was significantly reduced to 1.5 ± 0.3, reaching the lower limit of detection. ハムスター hamsters treated with hIg or NIP in hamster plasma ハムスター Alveolar epithelial cell hyperplasia and intraalveolar eosinophils were not observed in any of the hamsters treated with IFN-γ (Fig. 2f, data not shown).

[0208] Consideration We investigated CCP as an intranasal prophylactic agent in a preclinical hamster model. Our study demonstrated a reduction in infectious virus in 78% of treated hamsters, with no detectable virus in half of them. A further reduction in infectious virus was observed in all hamsters after intranasal administration of convalescent plasma from vaccine recipients. This is due to the significantly increased neutralization of SARS-CoV-2 infection after vaccination. Importantly, intranasal administration of human-donated CCP to exposed hamsters resulted in a type of lung tissue injury distinct from the typical infectious inflammation caused by SARS-CoV-2. Instead, alveolar epithelial cell hyperplasia and intraalveolar eosinophils suggested allergic toxicity. This characteristic histopathological picture was not observed in any hamsters treated with hamster CCP, indicating a cross-species side effect.

[0209] Intranasal administration of monoclonal antibodies has demonstrated clinical potential for preventing COVID-19 in preclinical animal studies, however, the recombinant anti-SARS-CoV-2 IgG1 monoclonal antibody tested for prophylaxis in our infection model did not prevent or reduce viral load.

[0210] Furthermore, all CCP samples used in our study had very low anti-SARS-CoV-2 immunoglobulin content per dose: 0.01 mg / kg for CCP, 0.05 mg / kg for purified hIg in buffer, and 0.06 mg / kg for VCCP. Consequently, intranasal CCP prevents or reduces SARS-CoV-2 infection at ultra-low doses.

[0211] Furthermore, treatment with VCCPs resulted in reduced viral infectivity compared to purified hIgs in buffer, an additional advantage of VCCPs that may be due to the plasma matrix and / or high levels of IgA.

[0212] Our results suggest that intranasal administration of CCP to humans during the critical early phase of epigenetic or pandemic responses could protect a significant portion of the population. If an oropharyngeal CCP spray were as effective in vulnerable humans as in our in vivo animal studies, it would prevent 40% of infections and slow disease progression in the remaining 40%. Further development is envisioned for an oropharyngeal spray to protect the elderly, immunocompromised patients, and healthcare workers. CCP-loaded oropharyngeal sprays could also protect populations in low- and middle-income countries (LMICs) with limited primary access to vaccines or in countries where vaccine skepticism is strong. In the general population, such a CCP spray could help limit the spread of disease, even among vaccinated individuals who are currently most protected from hospitalization but not infection.

[0213] Because the availability of CCP is crucial for ultimately providing the potential (oral) throat spray (sub)population, we can use figures from Belgium to estimate the balance between supply and demand. In Belgium, the first wave began in March 2020. By the end of April, 5% of Belgian donors tested positive for SARS-CoV-2 antibodies, corresponding to 12,500 (known) donors. Four months later, a second wave hit Belgium, resulting in 7,500 hospitalizations in a short period of time and placing a significant strain on the healthcare system. Assuming that all of these hospitalized COVID-19 cases were "vulnerable" in advance and that this population could have been identified in May based on medical records, 1,200 liters of CCP would be required to supply them with twice-daily (200 μL / nostril) doses of CCP (oral) throat spray for 200 consecutive days (i.e., winter) in September. This simply means that the CCP successfully completed 2,400 donations since May. This would have been feasible with 12,500 potential donors from the first wave and a logistics system capable of handling 200,000 plasma donations per year.

[0214] Future epidemics and pandemics will be best addressed with preventative medicines such as vaccines to prevent people from becoming sick at home or in hospitals. Our studies suggest that convalescent plasma could be developed in a similar way for SARS-CoV-2 and possibly other airborne pathogens.

[0215] Example 2. Humoral immunity after intranasal administration of convalescent plasma In this experiment, we aim to follow the humoral immunity against viral infection in hamsters treated intranasally with COVID-19 convalescent plasma (CCP). We aim to measure the impact on humoral responses in hamsters with reduced viral loads after intranasal CCP treatment. The functionality of the immune response will be assessed by (a) the presence of IgG-type antiviral antibodies 21 days after primary infection and (b) reinfection of hamsters with the same virus strain as the primary infection (but at a lower dose).

[0216] method Sentinel hamsters are treated intranasally with hamster CCP (test group) or naive hamster plasma (control group) for five consecutive days. The sentinels are co-housed with SARS-CoV-2-infected index hamsters (transmission model) for four days. Infection and treatment begin one day before co-housing. Viral load is monitored by RNA using RT-PCR and infectious virus by TCID on pharyngeal mucosa-transmitted swabs. Clinical disease progression in the lungs is measured by CT scans at the estimated peak of infection in the hamsters (day 4 for sentinels and day 3 for index hamsters). Humoral immunity is assessed by ELISA in serial blood samples collected starting on day 6 after infection or treatment. The humoral response is followed until day 28. On this day, the hamsters are reinfected with virus to assess the functionality of the immune response generated in the sentinel hamsters. As a control, the index hamster is followed, including a non-immunized control hamster, to verify that the second infection was successful. Viral loads were measured daily for 4 days, and blood samples were taken every other day. On day 32, CT scans were performed to assess the clinical status of the lungs after reinfection. The hamsters were then sacrificed, and lungs were harvested for histological evaluation, infectious load, and viral RNA measurement.

Claims

1. 1. A composition for use in preventing infection and / or transmission by a causative agent that invades or spreads in the nasopharyngeal region of a subject, or in preventing or treating a disease caused by said causative agent, the composition comprising plasma and immunoglobulins against said causative agent, the immunoglobulins being obtained from a donor subject that has developed a humoral immune response to said causative agent, and the composition being administered intranasally or to the nasopharyngeal region.

2. 2. The composition for use of claim 1, wherein the causative agent causes a respiratory infection, the composition comprises plasma and immunoglobulins against the causative agent of the respiratory infection, the immunoglobulins are obtained from a donor subject who has developed a humoral immune response against the causative agent of the respiratory infection, and the composition is administered intranasally or to the nasopharyngeal region.

3. 3. The composition for use according to claim 1 or 2, which is used to prevent or reduce the progression of the infection to a lower respiratory, systemic or gastrointestinal infection and / or to prevent the transmission of the infection within a population.

4. 4. The composition for use according to any one of claims 1 to 3, wherein the composition comprises at least 70% (v / v) of the plasma and immunoglobulins against the causative agent of the infectious disease, the immunoglobulins being obtained from a donor subject who has developed a humoral immune response against the causative agent of the infectious disease.

5. The composition for use according to any one of claims 1 to 4, wherein the plasma is not rich in platelets, preferably the plasma is not platelet rich plasma (PRP).

6. 5. The composition for use according to any one of claims 1 to 4, wherein the composition comprises convalescent plasma obtained from a donor subject who has developed a humoral immune response against the causative agent of the infectious disease.

7. The composition comprises: - before the subject is exposed to the agent that causes the infectious disease; - before the subject becomes infected with the causative agent of the infectious disease; - after the subject has been exposed to the agent causing the infectious disease; or - after the subject has been infected with the infectious agent, A composition for use according to any one of claims 1 to 6, administered to said subject.

8. The composition for use according to any one of claims 1 to 7, wherein the causative agent of the infectious disease is a virus, bacterium, bacterial spore or fungus, preferably a virus.

9. The composition for use according to any one of claims 1 to 8, wherein the respiratory infection is a respiratory infection.

10. 10. The composition for use according to claim 9, wherein the respiratory infection is a viral respiratory infection, preferably the causative agent of the viral respiratory infection is selected from the group consisting of respiratory syncytial virus (RSV), influenza virus, and coronavirus, preferably the respiratory virus is a coronavirus.

11. 11. The composition for use according to claim 10, wherein the coronavirus is a severe acute respiratory syndrome-associated coronavirus, most preferably severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2; causing COVID-19).

12. The composition for use according to any one of claims 1 to 11, wherein the composition is a pulmonary aerosol formulation, nasal drops, oral nasal drops, nasal spray, oral nasal spray, or nasopharyngeal spray.

13. 13. The composition for use according to any one of claims 1 to 12, wherein the composition is used as an adjunct to conventional infectious disease treatment, prevention or vaccination methods or is used in the absence of conventional infectious disease treatment, prevention or vaccination methods.

14. The composition for use according to any one of claims 1 to 12, wherein said composition is used in the absence of conventional methods of treating, preventing or vaccinating against infectious diseases.

15. A nasal, nasopharyngeal or oronasal spray comprising a composition comprising plasma and immunoglobulins against the causative agent of said infection, said immunoglobulins being obtained from a donor subject who has developed a humoral immune response against the causative agent of said respiratory infection as defined in any one of claims 1 to 14.

16. 15. A nasal, nasopharyngeal or oronasal spray device comprising a composition comprising plasma and immunoglobulins against a causative agent of said respiratory infection, said immunoglobulins being obtained from a donor subject who has developed a humoral immune response against said causative agent of said respiratory infection as defined in any one of claims 1 to 14.

17. The composition for use according to any one of claims 1 to 14, wherein said composition induces a humoral immune response against said causative agent in a subject to which said composition is administered.

18. A method for inducing a humoral immune response against a pathogen that invades or spreads in the nasopharyngeal region of a subject in need thereof, comprising administering to the subject a composition comprising plasma and immunoglobulins against the pathogen, wherein the immunoglobulins are obtained from a donor subject that has developed a humoral immune response against the pathogen, and the composition is administered intranasally or to the nasopharyngeal region.

19. 1. A method for producing a vaccine comprising combining plasma and a composition comprising immunoglobulins against a pathogen that invades or spreads in the nasopharyngeal region with one or more pharmaceutically acceptable excipients, wherein the immunoglobulins are obtained from a donor subject that has developed a humoral immune response against the pathogen.

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