Biological therapeutic products for infectious related diseases or conditions

Abstract

Disclosed are products and methods of use of the products for the prevention and treatment of infectious diseases and diseases or conditions induced by harmful antibodies. The harmful antibodies are induced during infection, or vaccination, or use of therapeutic antibodies. The products provided include immunoglobulin products, serum or plasma, specific antibodies against viral pathogens.

Description

[0001] This invention is a divisional application of application number CN201480015768.4 (PCT / US2014 / 025926), filed on March 13, 2014, entitled "Biotherapy Product for Infection-Related Diseases or Symptoms".

[0002] Priority requirements

[0003] This application claims priority to U.S. Provisional Patent Application No. 61 / 792,888, filed March 15, 2013, entitled "Biotherapy Product for Infection-Related Diseases or Conditions". The entire specification and disclosure of that application are incorporated herein by reference. Technical Field

[0004] This invention relates primarily to the fields of medicine and epidemiology, and in particular to products and methods for treating or preventing infections caused by viral pathogens, diseases and conditions caused by harmful antibodies induced by infectious pathogens, vaccines, and therapeutic antibodies. Background Technology

[0005] Infectious diseases are clinically recognized illnesses in humans or animals. Information on global mortality collected by the World Health Organization (WHO) indicates that infectious diseases account for up to 25.9% of all deaths worldwide, or 14,700,000 in 2002 (WHO World Health Report 2002). Current death tolls are even higher.

[0006] Influenza virus infections, especially influenza pandemics, threaten human health and the global economy. There is close attention focused on a potential influenza pandemic caused by the highly pathogenic H5N1 (avian) influenza virus, which has caused far more deaths than the 2009 swine flu pandemic (WHO). Currently, there are no effective treatments for severe illness caused by influenza virus infections, particularly for severe illness occurring more than 48 hours after infection.

[0007] Vaccines are the most effective way to prevent infectious diseases. However, vaccines are not perfect, as they can cause serious adverse reactions and even death. For example, the 1976 swine flu vaccine may have been associated with approximately 500 cases of Guillain-Barré syndrome (GBS) and 25 deaths, leading to a recall of the vaccine (US CDC, VAERS). In 2009, a monovalent H1N1 (swine) flu vaccine may have caused 636 serious health events, including 103 cases of GBS and 51 deaths in the United States (US CDC, VAERS). To date, there is no direct evidence linking influenza vaccines to serious adverse reactions. Because the pathogenesis is not yet clear, there are no drugs available to prevent or treat serious adverse reactions caused by influenza vaccines or other vaccines.

[0008] An estimated 2,640,000 infants die each year during gestation at 28 weeks or longer (stillbirth) (Tien adv. V., et al. Major risk factors for stillbirth in high-income countries: a systematic review and meta-analysis. Lancet, April 16, 2011; 377:1331-1340), and 4,000,000 infants die within the first four weeks after birth (neonatal death) (Lawn JE, Cousens S, Zupan J. 4 million Neonatal deaths: When? Where? Why? Lancet, 2005 Mar 5-1l; 365(9462):891-900). One potential risk factor may be related to infection. However, it is unclear how infection induces fetal or neonatal death.

[0009] We have developed virus-free animal models by injecting various antipathogenic antibodies into chicken embryos or pregnant mice. Experimental results using these animal models indicate that some antipathogenic antibodies induced during infection (such as influenza virus infection) or through vaccines (such as influenza vaccines) can be harmful and may lead to severe illnesses such as GBS or even death. Therapeutic products for the prevention and treatment of diseases caused by these toxic antibodies have been tested using these animal models. Summary of the Invention

[0010] This invention discloses products for the prevention and treatment of infectious diseases and diseases or conditions induced by harmful antibodies, as well as methods of using the products. The harmful antibodies are induced during infection, vaccination, or the use of therapeutic antibodies. The products provided by this invention include immunoglobulin products, serum or plasma, and specific antibodies against viral pathogens. Attached Figure Description

[0011] Figure 1 The image shows histological changes in the lungs of suckling mice born to mothers treated with human serum from healthy individuals (A) or serum against the 09H1N1 influenza virus (B). These data suggest that antibodies induced by the 09H1N1 influenza virus are harmful to the fetus and newborn, as they can lead to late fetal death (stillbirth) and neonatal death. Detailed Implementation

[0012] This invention can be implemented in many different ways, and preferred and alternative embodiments will be described in detail herein. However, it should be understood that the disclosure of this invention should be considered as an example of inventive principles and is not intended to limit the spirit and scope of the invention and / or the claims of the illustrated embodiments.

[0013] Dual action of antibodies

[0014] Based on traditional understanding, antibodies induced by infectious pathogens or vaccines provide protection to the host because they neutralize pathogens and prevent or treat infectious diseases. However, the effect of antibodies is bidirectional. Some antibodies can cross-react with certain cells, tissues, or organs of the host, thereby triggering harmful reactions such as antibody-dependent cytotoxicity or defects in signal transduction pathways, and causing tissue and organ damage or disease. For example, antiviral antibodies can bind to host tissues and organs, stimulating and causing disease in those tissues and organs (e.g., autoimmune diseases as described in PCT / US2007 / 018258 and PCT / US2009 / 039810).

[0015] It is known that in acute viral infections or vaccinations, antibodies against pathogens (such as viruses) or vaccines are induced and elevated one week after infection or vaccination, peaking two weeks after infection or vaccination. Some antibodies can bind to certain types of host cells, tissues, or organs. This process can be pathogenic and can cause severe illness or symptoms, or even death. For example, anti-rotavirus antibodies bind to proliferating goblet cells during rotavirus infection (PCT / US2009 / 039810 and US201 10085981). Injection of high doses of anti-rotavirus antibodies into pregnant mice can induce death in pups born to the dames and proliferation (inflammation) of bile duct epithelial cells (PCT / US2009 / 039810, US201 10085981). Furthermore, as described in PCT / US2009 / 039810 and US20110085981, administration of high doses of anti-rotavirus antibodies to suckling mice before or after rotavirus infection can cause death or severe infection in the suckling mice.

[0016] In the present invention, the term "harmful antibody" refers to any antibody capable of causing pathological reactions, damage, or disease in the host's cells, tissues, and organs. Such harmful antibodies may be induced during infection (e.g., influenza virus infection) or vaccination (e.g., influenza vaccination), or passively introduced (e.g., therapeutic antibodies). Diseases or conditions caused by the harmful antibodies of this invention include, but are not limited to, infectious diseases, serious adverse reactions to vaccines or therapeutic antibodies, infection-related autoimmune diseases, allergies, inflammation, infection-related tumors, and any other conditions (known or unknown) induced by harmful antibodies.

[0017] Animal models used for pathological research and evaluation of the safety of vaccines and antibodies

[0018] One embodiment of the present invention discloses an experimental model established by injecting antipathogenic antibodies into chicken embryos, pregnant mice, or newborn suckling mice. In one aspect, the present invention provides an experimental model developed by injecting human anti-influenza immune serum or other antipathogenic antibodies into chicken embryos, and observing the health status of newborn chicks as described in the examples. The human immune serum is obtained from patients infected with natural influenza viruses or vaccinated against influenza, or the human immune serum is derived from other infectious diseases. As described in the examples, injection of human anti-influenza virus serum into 16-day (E16) and E19-day chicken embryos induced leg deformities in newborn chicks, similar to Guillain-Barre syndrome (GBS) in humans. The frequency of GBS-like symptoms induced by anti-influenza virus antibodies is listed in Table 1. Results from this animal model indicate that antibodies induced by the 2009 H1M (swine) virus have the highest risk of inducing GBS (42.5%), followed by antibodies induced by influenza B virus (32.5%), avian influenza H5N1 virus (27.5%), seasonal H1N1 virus (17.5%), and H3N2 virus (15.0%). Furthermore, injection of human anti-influenza virus serum can lead to fetal and neonatal death. Mortality rates are listed in Table 2. Results from this animal model indicate that antibodies induced by influenza B virus have the highest risk of causing fetal and neonatal death (65.0%), followed by antibodies induced by avian influenza H5N1 virus (57.5%), 2009 H1N1 (swine) virus (52.5%), H3N2 virus (37.5%), and seasonal H1N1 virus (15.0%). As a control, anti-respiratory syncytial virus (RSV) serum did not cause significant GBS-like symptoms or death (Tables 1-2).

[0019] This experimental model can be used, but is not limited to, for the study of infectious diseases, autoimmune diseases such as Guillain-Barre syndrome (GBS) or GBS-like conditions, infection-related fetal and neonatal deaths, pathological studies of adverse reactions to influenza vaccines and other vaccines or therapeutic antibodies; for screening drugs (those described above) for the prevention and treatment of conditions or illnesses caused by harmful antibodies; or for the rapid evaluation of the safety of vaccines (e.g., influenza vaccines) and therapeutic antibodies. Other embodiments besides those described above are readily apparent. Various other aspects, objectives, features, and advantages will become apparent from the detailed description of the invention.

[0020] In another aspect, the present invention discloses an experimental model developed by injecting anti-influenza immune serum or other anti-pathogen antibodies into pregnant mice and observing the health status of pregnant mother mice and newborn suckling mice as described in the examples. As described in the examples, injection of human anti-influenza virus serum into pregnant mice at 16 days of gestation (E16) and E19 can cause abortion, premature birth, or prolonged labor in the pregnant mice, as well as fetal and neonatal death in newborn suckling mice delivered to the dame by these mother mice. The frequency of fetal and neonatal death in newborn suckling mice is listed in Table 3. The results using this animal model indicate that antibodies induced by H5N1 (avian influenza) virus have the highest risk of death (34.6%), followed by antibodies induced by seasonal H1N1 virus (26.7%), H3N2 virus (26.5%), influenza B virus (23.5%), and 2009 H1N1 (swine) virus (22.9%). The abortion frequency in pregnant mice is listed in Table 4. Results using this animal model indicate that antibodies induced by influenza B virus had the highest risk of causing abortion (33.3%), followed by antibodies induced by 2009 H1N1 (swine) virus (31.3%), H3N2 virus (28.6%), seasonal H1N1 virus (21.4%), and H5N1 (avian influenza) virus (20.0%). Histological changes in dead mice showed erythrocyte infiltration, inflammatory cell and epithelial cell proliferation, and localized tissue lesions in the lungs, brain, heart, and liver of dead chickens and suckling mice. Furthermore, human IgG bound to these organs could be detected in tissue sections from dead chickens and suckling mice.

[0021] In summary, the data from the aforementioned animal models strongly suggest that influenza virus infection or vaccination in pregnant female animals may be harmful to the fetus and newborn, as some antibodies induced by the influenza vaccine may lead to miscarriage, premature birth, prolonged labor, and fetal and neonatal death. Furthermore, some antibodies induced by the influenza virus or influenza vaccine may cause autoimmune diseases or serious adverse reactions, such as GBS or GBS-like symptoms and death. Similarly, some antibodies induced by other pathogens or other vaccines may also cause autoimmune diseases or serious adverse reactions.

[0022] This experimental model can be used, but is not limited to, pathological studies of infectious diseases, autoimmune diseases, infection-related fetal and neonatal deaths, influenza vaccines, and adverse reactions to other vaccines or therapeutic antibodies; screening for drugs (as described above) for the prevention and treatment of diseases or conditions caused by harmful antibodies; or rapidly evaluating the safety of vaccines (e.g., influenza vaccines) and therapeutic antibodies. Other embodiments besides those described above are readily apparent. Various other objectives, features, and advantages of the invention will become apparent from the detailed description of the experimental model.

[0023] Novel pathological mechanisms of infection and infection-related diseases or conditions

[0024] One embodiment of the present invention discloses a novel pathological mechanism of infection-related diseases or conditions, including but not limited to infectious diseases, autoimmune diseases, cancer, and other diseases that may be related to infection.

[0025] On the one hand, this invention discloses a novel concept of the pathological mechanism of infection. This mechanism includes, but is not limited to: 1) initial, primary damage to cells, tissues, or organs caused by a virus within one week; 2) secondary damage caused by antiviral antibodies, which begin to increase in the first week after viral clearance and reach peak levels in the second to third weeks; 3) the secondary damage can further superimpose on the primary damage and cause severe illness, even death (primary damage plus secondary damage); 4) the secondary damage lasts longer and has a wider impact because the antibodies persist longer than the virus. The primary damage plus secondary damage described above can cause severe illness during acute infection; and persistently persisting antibodies can cause or aggravate diseases or conditions, such as autoimmune diseases, allergies, cancer, and other diseases that may be associated with toxic antibodies.

[0026] Based on the aforementioned pathological mechanisms, antibodies against the viral strains of the 1918 and 2009 influenza pandemics may have been the leading cause of death during those pandemics. This is supported by the following evidence: 1) Approximately two-thirds of deaths caused by influenza virus infection occurred during the second and third weeks (Edwin O. Jordan. Epidemic Influenza - A survey, 1927), which coincides with the time when antibody levels reached their peak; and 2) Most severe or fatal infections occurred in young adults (Edwin O. Jordan. Epidemic Influenza - A survey, 1927; WHO: Pandemic (H IM) 2009 - update 41), because younger individuals can induce higher levels of antibodies.

[0027] On the other hand, this invention discloses novel pathological mechanisms and the etiologies of autoimmune diseases. Based on the above observations, some antibodies induced by pathogens or vaccines can bind to the body's own cells, tissues, or organs and mislead the immune response, leading to self-attack. One example is that antibodies induced by influenza virus or influenza vaccine can cause GBS or fetal and neonatal death as described above. Another example is that injecting high doses of anti-rotavirus antibodies into pregnant mice can lead to the death of sucklings born to the mother and bile duct obstruction (biliary atresia) caused by epithelial cell proliferation (inflammation) (PCT / US2009 / 039810, US201 10085981).

[0028] On the other hand, this invention also discloses novel pathological mechanisms of cancer or other diseases (known or unknown). Many other disease processes, such as cancer, diabetes, heart disease, and other diseases (known or unknown), are caused by uncontrolled cell growth or defects in signal transduction pathways. Certain antibodies or therapeutic antibodies induced during infection or vaccination can bind to certain cell surface molecules and induce cell proliferation or defects in signal transduction pathways. Repeated or persistent stimulation by such harmful antibodies can lead to uncontrolled cell proliferation or defects in signal transduction pathways, which can lead to the occurrence and development of various cancers, autoimmune diseases, or other conditions (known or unknown). For example, anti-HIV antibodies stimulate the proliferation of human lung cancer cells (A549) and liver cancer cells (HepG-2). Some harmful antibodies induced during acute or chronic infections (such as human T-cell leukemia virus infection) can cause leukemia or lymphoma. These harmful antibodies can bind to leukemia cells, including lymphocytes, and stimulate these blood cells to undergo uncontrolled cell proliferation, thereby leading to leukemia or lymphoma.

[0029] Other embodiments besides those described above are readily apparent. Various other objectives, features, and advantages of the present invention are also readily apparent based on novel etiologies (harmful antibodies) and novel pathological mechanisms.

[0030] Diseases and symptoms induced by harmful antibodies

[0031] According to the present invention, harmful antibodies can be induced during infection (such as influenza virus infection) or vaccination (such as influenza vaccination), or passively introduced (such as therapeutic antibodies). Diseases or conditions caused by the harmful antibodies disclosed in this invention include, but are not limited to, infectious diseases, serious adverse reactions caused by vaccines or therapeutic antibodies, infection-related autoimmune diseases, allergies, and inflammation, as well as infection-related tumors and other diseases (known or unknown).

[0032] The term "infectious disease" refers to the invasion of a host organism's tissues by a disease-inducing organism and its replicas, and the host organism's response to these organisms and the toxins they produce. Short-term infections are acute infections. Long-term infections are chronic infections. Specific pathogens applicable to this process include, but are not limited to, viruses, bacteria, parasites, fungi, viroids, prions, protozoa, and insects. Types of pathogens include, but are not limited to, live or dead or inactivated pathogens, fresh or dried pathogens, fixed or frozen pathogens, whole or partial or fragmented pathogens, sections, smears, homogenates, lysates, and extracts of pathogens of any type. Examples of infection include, but are not limited to, those caused by influenza virus, respiratory enterovirus, rotavirus, cytomegalovirus (CMV), Epstein-Barr virus (EBV), adenovirus, hepatitis viruses including HAV, HBV, HCV, human immunodeficiency virus (HIV), human T-cell leukemia virus (HTLV), human papillomavirus (HPV), poliovirus, parainfluenza virus, measles virus, mumps virus, respiratory syncytial virus (RSV), human herpesvirus (HHV), herpes simplex virus (HSV), varicella-zoster virus, classical swine fever virus, poxvirus, rabies virus, canine distemper virus, foot-and-mouth disease virus, rhinovirus, Newcastle disease virus, pseudovirus. Rabies virus, cholera, syphilis, anthrax, leprosy, and the Black Death, rickettsia, gonococci, Bordetella pertussis, Escherichia coli, Salmonella, Vibrio cholerae, Pseudomonas aeruginosa, Yersinia pestis, Tulafrancella, Haemophilus influenzae, purple sulfur bacteria, Helicobacter pylori, Campylobacter jejuni, Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis, Clostridium tetani, Clostridium botulinum, Staphylococcus, Streptococcus pneumoniae, Diplococcus pneumoniae, Mycoplasma, Bacteroides fragilis, Mycobacterium tuberculosis, Mycobacterium leprae, Corynebacterium diphtheriae, Treponema pallidum, Borrelia burgdorferi, Chlamydia trachomatis, Chlamydia psittaci, phycocyanin, phycoerythrin, mitochondria, chloroplasts, etc., are not limited to diseases caused by these bacteria.

[0033] According to the present invention, the term "infection-related autoimmune disease, allergy, and inflammation" refers to a disease or condition caused by harmful antibodies induced during infection or vaccination, or harmful antibodies introduced by antibody therapy. This disease or condition typically develops some time after infection or vaccination (e.g., within 4-8 weeks). Examples of infection-related autoimmune diseases, allergies, and inflammation include, but are not limited to, Guillain-Barre syndrome, autism, Kawasaki's disease, biliary atresia, primary biliary cirrhosis, systemic lupus erythematosus, leukemia, acute leukemia, rheumatoid arthritis, adult-onset diabetes mellitus (type II diabetes), Sjogren's syndrome, juvenile diabetes mellitus, Hodgkin's and non-Hodgkin's lymphoma, malignant melanoma, cryoglobulinemia, and hepatitis B virus infection. Hepatitis C virus infection, Wegener's granulomatosis, inflammatory bowel disease, polymyositis, dermatomyositis, multiple endocrine disorders, Schmidt's syndrome, autoimmune uveitis, Addison's disease, adrenalitis, Graves' disease, thyroiditis, Hashimoto's thyroiditis, autoimmune thyroid disease, pernicious anemia, thyroid gland, gastric atrophy, chronic hepatitis, lupus-like hepatitis, atherosclerosis, Alzheimer's disease, demyelating disorders. Diseases, multiple sclerosis, subacute cutaneous lupus erythematosus, hypoparathyroidism, Dressler's syndrome, myasthenia gravis, autoimmune thrombocytopenic purpura, idiopathic thrombocytopenic purpura, hemolytic anemia, autoimmune hemolytic anemia, herpetic dermatitis, alopecia areata, autoimmune cystitis, bullous pemphigoid, scleroderma, progressive systemic sclerosis, CREST syndrome (calcification, Raynaud's esophageal motility disorder, fingertip sclerosis, telangiectasia), male or female Autoimmune infertility, ankylosing spondylitis, ulcerative colitis, Crohn's disease, mixed connective tissue disease, polyarteritis nodosa, systemic necrotizing vasculitis, juvenile rheumatoid arthritis, nephritis, allergic dermatitis, allergic rhinitis, Goodpasture's syndrome, Chagas' disease, sarcoidosis, rheumatic fever, asthma, recurrent miscarriage, antiphospholipid syndrome, farmer's lung disease, erythema multiforme, pemphigus vulgaris, pemphigus, bullous pemphigoid, postcardotomy syndrome, Cushing's syndrome, autoimmune chronic active hepatitis, bird lover's lung disease, asthma, allergic diseases, allergic encephalomyelitis, toxic necrodermal lysis, alopecia, Alportt's syndrome, alveolar alveolitis, allergic alveolar alveolitis, fibrotic alveolar alveolitis, interstitial lung disease, erythema nodosa, pyoderma gangrenosa.Transfusion reactions, leprosy, malaria, leishmaniasis, trypanosomiasis, chronic fatigue syndrome, fibromyalgia, Takayasu arteritis, polymyalgia rheumatica, temporal arteritis, schistosomiasis, giant cell arteritis, ascariasis, aspergillosis, Sampter's syndrome (also known as trisomy, nasal polyps, eosinophilia, and asthma), Behcet's disease, Caplan's syndrome, dengue fever, encephalomyelitis, endocarditis, myocarditis, endocardial fibrosis, endophthalmitis, erythema elevatum Psoriasis, fetal nucleocytosis, eosinophilic fasciitis, Shulman's syndrome, Felty's syndrome, filariasis, cyclitis, chronic cyclitis, heterochromia iridis, Fuch's cyclitis, IgA nephropathy, Henoch-Schönlein purpura, glomerulonephritis, graft-versus-host disease, transplant rejection, cardiomyopathy, Alzheimer's disease, parvovirus infection, rubella virus infection, post-vaccination syndrome, congenital rubella infection, renal cell carcinoma, multiple myeloma, Eaton-Lambert syndrome, relapsing polychondritis, Waldenstrom's macroglobulinemia, mumps virus infection, thrombotic thrombocytopenic purpura, and any other disease or condition in which the specific recognition of pathogen-induced or vaccine-induced antibodies is suspected or shown to play an important role in any aspect of the pathogenesis of the clinical disease.

[0034] According to the present invention, the term "serious adverse reaction caused by vaccine or therapeutic antibody" refers to a serious illness or condition caused by harmful antibodies induced during vaccination or antibody treatment. This illness or condition typically develops some time after vaccination or antibody treatment (e.g., within 4-8 weeks). Examples of serious adverse reactions caused by vaccines disclosed in this invention include, but are not limited to, death, sudden infant death syndrome, Guillain-Barre syndrome, Kawasaki's disease, acute leukemia, allergy, severe allergic reaction, asthma, epilepsy, immune system disorders, behavioral disorders, neurological damage, permanent brain damage, learning difficulties, convulsions, severe convulsions, decreased consciousness, autism, prolonged coma, headache, upper and lower respiratory tract infections, arthralgia, abdominal pain, cough, nausea, diarrhea, fever, hematuria or bloody stools, pneumonia, gastritis or enteritis, incessant crying, fainting, deafness, transient thrombocytopenia, urticaria, joint pain, intussusception, vomiting, severe neurological reactions, life-threatening serious illnesses with organ failure characteristics, stillbirth, neonatal death, and any other disease or condition in which the specific recognition of pathogen-induced or vaccine-induced antibodies is suspected or shown to play an important role in any aspect of the pathogenesis of the clinical disease.

[0035] According to the present invention, the term "infection-associated tumor" refers to a disease or condition characterized by uncontrolled cell growth caused by harmful antibodies induced during acute infection, vaccination, or chronic infection, or antibody therapy. This disease or condition typically develops some time after infection, vaccination, or antibody therapy (e.g., acute leukemia). Additionally, many tumors arise from repeated stimulation by antibodies induced during chronic infection. Examples of infection-related tumors or cancers disclosed in this invention include, but are not limited to, acute or chronic leukemia, Hodgkin's and non-Hodgkin's leukemia, malignant melanoma, tumors or cancers occurring during HIV, HTLV, HBV, HCV, EBV, CMV, HPV, HHV, HSV, adenovirus, measles virus, mumps virus, varicella-zoster virus infection, lung cancer, liver cancer, prostate cancer, breast cancer, colon cancer, gastrointestinal cancer, pancreatic cancer, cervical cancer, all cancers, thyroid cancer, neurotumor or tumor, renal cell carcinoma, kidney cancer, multiple myeloma, hemangioma, cardiac fibroma, and any other disease or condition in which the specific recognition of pathogen-induced or vaccine-induced antibodies is suspected or shown to play an important role in any aspect of the pathogenesis of the clinical disease.

[0036] Products for the treatment of conditions or illnesses induced by harmful antibodies.

[0037] Based on the novel etiologies and pathological mechanisms of diseases or symptoms induced by harmful antibodies as described above, one embodiment of the present invention discloses therapeutic means, substances, or products (new or existing) for preventing and treating diseases and symptoms caused by harmful antibodies by blocking the binding of harmful antibodies to host cells, tissues, and organs, or by neutralizing harmful antibodies, as well as the manner and method of using such substances or products (new or existing). The products provided by the present invention include, but are not limited to, serum or plasma, immunoglobulins, non-harmful antibodies, pathogen-derived products, host-derived products, polysaccharides, and any other substances or products (new or existing) capable of competing with or neutralizing harmful antibodies.

[0038] Immunoglobulins, serum, or plasma used for conditions caused by harmful antibodies.

[0039] One embodiment of the present invention discloses an immunoglobulin product, serum, or plasma, and a method of using the immunoglobulin product, serum, or plasma in the prevention and treatment of diseases and conditions associated with harmful antibodies resulting from infection, vaccination, or antibody therapy. The immunoglobulins in the immunoglobulin, serum, or plasma can competitively dilute and block the binding of harmful antibodies to host cells, tissues, or organs, and protect them from damage by harmful antibodies.

[0040] On the one hand, the present invention discloses immunoglobulin products, serum or plasma for the prevention and treatment of diseases or conditions (such as fetal and neonatal death) caused by harmful antibodies.

[0041] The amount or concentration of the immunoglobulin product is from about 0.001 g to about 100 g. The amount of serum or plasma is from about 0.1 ml to 1000 ml. The immunoglobulin product, serum, or plasma is in the form of a solution, lyophilized form, injectable form, infusion form, or conjugated to nanoparticles, or other forms of use known to those skilled in the art.

[0042] The immunoglobulin product, serum, or plasma is used to prevent or treat diseases or conditions caused by the harmful antibodies disclosed in this invention. These diseases or conditions include, but are not limited to, infectious diseases, severe adverse reactions caused by vaccines or therapeutic antibodies, infection-related autoimmune diseases, allergies, inflammation, infection-related tumors, and other diseases (known or unknown) as described above.

[0043] Another embodiment of the present invention discloses a method for using immunoglobulin products or serum or plasma for the prevention and treatment of diseases or conditions caused by harmful antibodies disclosed in the present invention. Such diseases or conditions include, but are not limited to, infectious diseases, severe adverse reactions caused by vaccines or therapeutic antibodies, infection-related autoimmune diseases, allergies, inflammation, and infection-related tumors, as well as other diseases (known or unknown) as described above. One aspect of the method includes administering immunoglobulin products or serum or plasma to a human or animal individual suffering from or at risk of developing a disease or condition induced by harmful antibodies as described above.

[0044] Another aspect of this method includes administering immunoglobulin products or serum or plasma to a lactating animal or human mother whose infant has or is at risk of developing a disease or condition induced by harmful antibodies as described above.

[0045] The pharmaceutical composition or therapeutic product can be delivered to a biological organism, including a lactating mother as described above, via a variety of routes, including, for example, subcutaneous injection, open or closed external application, oral administration, intramuscular injection, intravenous injection (boost and infusion), intraperitoneal injection, intracavitary administration, or transdermal administration, inhalation administration, or other forms of use known to those skilled in the art of pharmaceutical manufacturing.

[0046] For example, as described in the examples, injecting human immunoglobulin or healthy human serum or plasma into suckling mice before or after infection with highly pathogenic influenza virus can reduce the mortality rate of infected mice by approximately 50% (Table 7).

[0047] On the other hand, the present invention discloses immunoglobulin products, serum, or plasma, and methods for using the immunoglobulin products, serum, or plasma to prevent and treat serious adverse reactions caused by antibodies induced by vaccines. For example, as described in the embodiments, the injection of human immunoglobulin, human serum, or human plasma into pregnant mice prevented fetal and neonatal death (Tables 5-6), miscarriage, premature birth, or prolonged labor in the mother mice caused by antibodies induced by influenza vaccines.

[0048] On the other hand, the present invention discloses immunoglobulin products or serum or plasma, and methods for using the immunoglobulin products or serum or plasma to prevent and treat fetal and neonatal death caused by infection or vaccine.

[0049] On the other hand, the present invention discloses a method for using human immunoglobulin products to prevent and treat infection-related autoimmune diseases, allergies and inflammation.

[0050] On the other hand, the present invention discloses a method for using human immunoglobulin products or human serum to prevent and treat infection-related inflammation.

[0051] For the therapeutic applications described above, the effective dose of serum or plasma is from about 0.1 ml / kg to 100 ml / kg; the effective dose of human Ig products is from about 1 mg / kg to 100 mg / kg.

[0052] Other features or characteristics of various therapeutic targets will become apparent from the detailed description of the invention.

[0053] A method for preparing better human immunoglobulin or antibody products

[0054] One embodiment of the present invention discloses a method for preparing better human immunoglobulins, the method comprising mixing an immunoglobulin product or a therapeutic antibody with a certain amount of N-acetylneuraminic acid, or an analogue of N-acetylneuraminic acid (such as N-acetylneuraminic acid methyl ester), or an analogue of N-acetylneuraminic acid plus N-acetylneuraminic acid (such as N-acetylneuraminic acid methyl ester).

[0055] The effective amount or effective concentration of N-acetylneuraminic acid or its analogues (such as N-acetylneuraminic acid methyl ester) is from 0.01 mg / g or 0.01 mg / ml to 100 mg / g or 100 mg / g.

[0056] Methods of using sialic acid in combination with human immunoglobulin or serum or plasma products

[0057] One embodiment of the present invention discloses a method for administering an immunoglobulin or serum or plasma product, the method comprising mixing an immunoglobulin product or therapeutic antibody with a certain amount of N-acetylneuraminic acid, or an analogue of N-acetylneuraminic acid (such as N-acetylneuraminic acid methyl ester) or an analogue of N-acetylneuraminic acid plus N-acetylneuraminic acid (such as N-acetylneuraminic acid methyl ester), and administering the resulting mixture to a subject.

[0058] For the therapeutic applications described above, the effective dose of serum or plasma is from about 0.1 ml / kg to 100 ml / kg; the effective dose of human Ig products is from about 0.1 mg / kg to 100 mg / kg. The effective dose of N-acetylneuraminic acid or an analogue of N-acetylneuraminic acid (such as N-acetylneuraminic acid methyl ester) is from 0.1 mg / kg to 100 mg / kg.

[0059] A method for preparing better human immunoglobulin or antibody products

[0060] One embodiment of the present invention discloses a method for preparing a better human immunoglobulin product during production, the method comprising mixing immunoglobulin or serum or plasma product or therapeutic antibody with a certain amount of N-acetylneuraminic acid, or an analogue of N-acetylneuraminic acid (such as N-acetylneuraminic acid methyl ester) or an analogue of N-acetylneuraminic acid plus N-acetylneuraminic acid (such as N-acetylneuraminic acid methyl ester).

[0061] The effective amount or effective concentration of N-acetylneuraminic acid or its analogues (such as N-acetylneuraminic acid methyl ester) is from 0.01 mg / g or 0.01 mg / ml to 100 mg / g or 100 mg / g.

[0062] Methods of use for pathogens or vaccines used to treat conditions induced by harmful antibodies.

[0063] Vaccines or inactivated pathogens can be used as therapeutic agents to treat serious infections caused by antibodies induced during infection. The likely mechanism of action of these therapeutic vaccines or inactivated pathogens is the neutralization of harmful antibodies.

[0064] Specific antibodies used to treat related diseases or conditions.

[0065] One embodiment of the present invention discloses a specific antibody against a first pathogen, which is used to prevent and treat infections caused by a second pathogen. The present invention also discloses a method of using the specific antibody product to prevent and treat infection-related diseases and conditions. The antibody product comprises at least one of an immunoglobulin molecule or an immunologically active portion of an immunoglobulin molecule.

[0066] One aspect of the present invention discloses specific antibodies against a first pathogen (such as HAV, HBV, or RSV) for the prevention and treatment of diseases or conditions caused by a second pathogen (such as influenza virus).

[0067] The amount or concentration of the antibody against the first pathogen is from about 0.001 g to about 100 g. The antibody is in solution form, lyophilized form, injectable form, infusion form, or conjugated to nanoparticles, or other forms of use known to those skilled in the art.

[0068] Specifically, antibodies against the primary pathogen are used to prevent or treat diseases or conditions caused by a secondary pathogen. These diseases or conditions include, but are not limited to, infectious diseases, serious adverse reactions to vaccines or therapeutic antibodies, infection-related autoimmune diseases, allergies, inflammation, infection-related tumors, and other conditions (known or unknown) as described above.

[0069] Another embodiment of the present invention discloses a method for treating a disease or condition caused by a second pathogen using a specific antibody against a first pathogen. The disease or condition includes, but is not limited to, infectious diseases, severe adverse reactions to vaccines or therapeutic antibodies, infection-related autoimmune diseases, allergies, inflammation, infection-related tumors, and other conditions (known or unknown) as described above.

[0070] On one hand, the method involves administering a specific antibody against the first pathogen to a human or animal individual who has or is at risk of developing a disease or condition caused by a second pathogen.

[0071] On the other hand, the method involves administering a specific antibody against a first pathogen to a lactating animal or human mother whose infant has or is at risk of developing a disease or condition caused by a second pathogen.

[0072] The pharmaceutical composition or therapeutic product can be delivered to a biological organism, including a lactating mother as described above, via a variety of routes, including, for example, subcutaneous injection, open or closed external application, oral administration, intramuscular injection, intravenous injection (boost and infusion), intraperitoneal injection, intracavitary administration, or transdermal administration, inhalation administration, or other forms of use known to those skilled in the art of pharmaceutical manufacturing.

[0073] For example, as described in the embodiments, injecting human anti-HAV, human anti-HBV, human anti-RSV, or rotavirus antibodies into suckling mice before or after infection with highly pathogenic influenza virus strains can reduce the mortality rate of infected mice by approximately 50% (Table 8). Other embodiments include, but are not limited to:

[0074] a. Antibodies against H1N1 virus used to prevent infection with H1N1, H3N2 or H5N1 viruses;

[0075] b. Antibodies against H3N2 virus used to prevent H1N1 and H3N2 virus infection;

[0076] c. Antibodies against H5N1 virus used to prevent infection with H1N1, H3N2 or H5N1 viruses;

[0077] d. Antibodies against respiratory syncytial virus used to prevent influenza virus and respiratory syncytial virus infection;

[0078] e. Antibodies against hepatitis A virus used to prevent influenza virus and hepatitis A virus infection;

[0079] f. Anti-adenovirus antibodies used to prevent influenza virus infection; or

[0080] g. Antibodies against rotavirus used to prevent influenza virus infection.

[0081] On the other hand, the present invention discloses a method for using specific antibodies against a first pathogen to prevent and treat infection-related autoimmune diseases, allergies and inflammation caused by a second pathogen.

[0082] On the other hand, the present invention discloses a method for preventing and treating infection-related inflammation caused by a second pathogen by using specific antibodies against a first pathogen.

[0083] As described above and as illustrated in the examples (Tables 5, 6, 7), low doses of the following immune sera or antibodies can be used as therapeutic agents for antibody prophylaxis and antibody therapy against influenza virus infection.

[0084] a. Antibodies against seasonal H1N1 viruses used to prevent infection with 2009 H1N1 (swine), other H1N1 and H5N1 influenza viruses;

[0085] b. Antibodies against 2009H1N1 (swine) virus used to prevent infection by other H1N1 and H5N1 influenza viruses;

[0086] c. Antibodies against H5N1 virus used to prevent H1N1 influenza virus infection.

[0087] For the therapeutic applications described above, the effective dose of the specific antibody against the primary pathogen is from about 0.001 ml / kg to 100 ml / kg.

[0088] Other features or characteristics of various therapeutic targets will become apparent from the detailed description of the invention.

[0089] In vivo evidence of novel mechanisms of action of vaccination and antibodies

[0090] One embodiment of the present invention discloses a novel mechanism of action for vaccines and antipathogen antibodies. Antibodies induced by infection or vaccines, or acquired through passive immunization, bind to and block at least one binding site of the pathogen.

[0091] Another experimental model is established by injecting antibodies against a first pathogen into animals, followed by infection with a second infectious pathogen as described in the examples. This experimental model can be used, but is not limited to, screening and identifying common binding sites of different pathogens, and screening and testing new vaccines made from the first pathogen that can prevent infection caused by the second pathogen. For example, vaccines against HAV, HBV, RSV, and rotavirus, as described in the examples, can be used to prevent influenza virus infection.

[0092] As described in the examples, an alternative experimental model was established by injecting antibodies against a first pathogen into animals, followed by infection with a second infectious pathogen. For example, five-day-old suckling mice were injected with antibodies against HAV, HBV, RSV, adenovirus, and rotavirus, respectively, and then infected with influenza A / PR / 8 / 34 (H1N1) virus the following day. Compared to suckling mice pretreated with saline, those pretreated with antibodies against HAV, HBV, RSV, adenovirus, and rotavirus were either not infected with influenza virus or had a mild infection. None of the antibodies reacted with influenza virus; therefore, these antibodies could not have functioned by neutralizing influenza virus. The only possible mechanism of action for these antibodies against different pathogens (such as HAV) is to block the shared viral binding sites of influenza virus.

[0093] This experiment not only discovered a novel mechanism for vaccines or pathogen-induced antibodies, but also identified a novel vaccine made from a first pathogen that can induce the production of antibodies against a second pathogen; or a novel specific antibody against a first pathogen that can prevent infection caused by a second pathogen. Many other features or characteristics of this novel therapeutic agent and its application will become apparent from the detailed description of the invention.

[0094] This experimental model can be used, but is not limited to, the screening and identification of shared binding sites among different pathogens, and the screening and testing of new vaccines made from a first pathogen that induce antibodies capable of preventing infection caused by a second pathogen. For example, vaccines against HAV, HBV, RSV, and rotavirus, as described in the examples, can be used to prevent influenza virus infection.

[0095] New vaccines for infectious diseases

[0096] One embodiment of the present invention discloses a vaccine containing a first pathogen that induces the production of antibodies capable of preventing infection caused by a second pathogen. The present invention also discloses a method for preventing and treating infection-related diseases and conditions caused by a second pathogen using a specific vaccine made from a first pathogen capable of inducing antibody production.

[0097] On the one hand, the present invention discloses a vaccine containing a first pathogen (such as HAV, HBV, or RSV) that induces the production of antibodies capable of preventing and treating diseases or symptoms caused by a second pathogen (such as influenza virus).

[0098] Vaccines containing a first pathogen are used to prevent or treat diseases or conditions caused by a second pathogen. These diseases or conditions include, but are not limited to, infectious diseases, serious adverse reactions to vaccines or therapeutic antibodies, infection-related autoimmune diseases, allergies, inflammation, infection-related tumors, and other conditions (known or unknown) as described above.

[0099] Another embodiment of the present invention discloses a method for treating a disease or condition caused by a second pathogen using a specific antibody against a first pathogen. The disease or condition includes, but is not limited to, infectious diseases, severe adverse reactions to vaccines or therapeutic antibodies, infection-related autoimmune diseases, allergies, inflammation, infection-related tumors, and other conditions (known or unknown) as described above.

[0100] One aspect of this method includes administering a vaccine containing a first pathogen to a human or animal individual who has or is at risk of developing a disease or condition caused by a second pathogen.

[0101] Another aspect of this method includes administering a vaccine containing a first pathogen to a lactating animal or human mother whose infant has or is at risk of developing a disease or condition caused by a second pathogen.

[0102] The pharmaceutical composition or vaccine can be delivered to biological organisms, including lactating mothers as described above, through a variety of routes, including, for example, oral administration, intramuscular injection, inhalation administration, or other forms of use known to those skilled in the art of pharmaceuticals.

[0103] New methods of using influenza vaccines

[0104] One embodiment of the present invention discloses a method of using an influenza vaccine. As described in the examples, influenza vaccine-induced anti-2009H1N1 (swine) and anti-H5N1 immune sera (pretreated with Neu5Ac) can effectively prevent and treat A / PR / 8 / 34 (H1N1) viral infection in suckling mice (Tables 6-7). These results provide evidence that two different influenza viruses (such as 2009H1N1 and H5N1 viruses) can share at least one binding site. Therefore, by using antibodies induced by either of these viruses to block the shared binding site, infection with the virus (such as H5N1 virus) can be prevented and treated not only by preventing and treating infection with that virus, but also by preventing and treating infection with the other virus (such as H1N1 virus).

[0105] Another embodiment of the present invention discloses a method for preventing infection by different viral strains (such as A / PR / 8 / 34H1N1 virus or H5N1 virus) using a vaccine containing one viral strain (such as 2009H1N1 swine influenza virus). In one aspect, based on shared binding sites, the present invention discloses the following novel vaccines:

[0106] a. Vaccines containing H1N1 virus strains for the prevention of infection with different viruses, such as H1N1 or H5N1; and

[0107] b. Vaccines containing the H5N1 virus used to prevent H1N1 influenza virus infection.

[0108] Other examples of the vaccine include, but are not limited to:

[0109] a. Vaccines containing the H1N1 influenza virus used to prevent infection with H3H2 or H5N1 viruses;

[0110] b. Vaccines containing the H3N2 influenza virus used to prevent H1N1 virus infection;

[0111] c. Vaccines containing the H5N1 influenza virus used to prevent infection with H1N1 or H3N2 viruses;

[0112] d. Vaccines containing respiratory syncytial virus used to prevent influenza virus infection;

[0113] e. Vaccines containing hepatitis A virus used to prevent influenza virus infection;

[0114] f. Adenovirus-containing vaccines used to prevent influenza virus infection; or

[0115] g. Rotavirus-containing vaccines used to prevent influenza virus infection.

[0116] On the other hand, the present invention discloses a method for using a vaccine containing a first pathogen to prevent and treat infection-related autoimmune diseases, allergies and inflammation caused by a second pathogen.

[0117] For the therapeutic applications described above, the effective dose of a vaccine containing the first pathogen is the amount that induces low to moderate levels of antibodies.

[0118] As will become apparent from the detailed description of the invention, many other features or characteristics of the therapeutic target will be readily apparent.

[0119] Another embodiment of the present invention discloses a vaccine or inactivated pathogen that can be used as a therapeutic product for treating severe infections caused by antibodies induced during infection. On the other hand, the present invention discloses a method of administering a therapeutic product containing a vaccine or inactivated pathogen to a patient to prevent or treat diseases or conditions caused by antibodies against the vaccine or pathogen induced during infection. A possible mechanism of action for this therapeutic vaccine or inactivated pathogen is the neutralization of harmful antibodies.

[0120] Methods of using sialic acid in combination with other therapeutic products

[0121] One embodiment of the present invention discloses a method for administering N-acetylneuraminic acid in combination with other therapeutic products, such as, but not limited to, immunoglobulins, serum or plasma products or antibody products, during administration. The method includes mixing the therapeutic product with a certain amount of N-acetylneuraminic acid, or an analogue of N-acetylneuraminic acid (such as N-acetylneuraminic acid methyl ester), or N-acetylneuraminic acid plus an analogue of N-acetylneuraminic acid (such as N-acetylneuraminic acid methyl ester), and administering the resulting mixture to a subject.

[0122] As described in the examples and shown in Tables 6-8, the combined use of N-acetylneuraminic acid with nonspecific human serum, or nonspecific human immunoglobulin, or specific anti-influenza virus antibodies, or specific antibodies against non-influenza viruses can significantly and effectively prevent H1N1 influenza virus infection. Data indicate that N-acetylneuraminic acid significantly enhances the anti-infective efficacy of nonspecific human serum, or nonspecific human immunoglobulin, or specific anti-influenza virus antibodies, or specific antibodies against pathogens different from influenza viruses, and reduces the toxicity of specific harmful antibodies (such as anti-influenza virus antibodies).

[0123] For example, as described in the examples (Tables 6 and 7), in newborn mice, immune sera premixed with N-acetylneuraminic acid against 2009H1N1 (pig), seasonal H1N1, and H5N1 viruses showed the best efficacy and lowest toxicity in preventing and treating influenza virus infection caused by A / PK78 / 34 (H1N1) virus.

[0124] As described in another embodiment (Table 5), injection of medium or high doses of antibodies against 2009H1N1 (swine), seasonal H1N1, H5N1 avian, and influenza B viruses into chicken embryos on days 16 and 19 induced leg deformities in chicks, a condition similar to Guillain-Barre syndrome (GBS) in humans. However, chicks injected with the same antibodies premixed with N-acetylneuraminic acid (Neu5Ac) did not develop GBS-like symptoms (Table 5).

[0125] As described in another embodiment (Tables 5-7), on days 16 and 19 of the embryonic period, newborn sucklings born to mother mice treated with medium to high doses of antibodies against 2009H1N1 (swine), seasonal H1N1, H5N1 avian and type B influenza viruses became ill or died; newborn sucklings injected with immune serum mixed with N-acetylneuraminic acid did not become ill.

[0126] For the therapeutic applications described above, the effective dose of serum or plasma is from about 0.1 ml / kg to 100 ml / kg; the effective dose of human Ig products is from about 0.1 mg / kg to 100 mg / kg. The effective dose of N-acetylneuraminic acid or an analogue of N-acetylneuraminic acid (such as N-acetylneuraminic acid methyl ester) is from 0.1 mg / kg to 100 mg / kg.

[0127] The possible mechanism of action of N-acetylneuraminic acid is to target harmful antibodies; therefore, it is effective when administered one week after vaccination or viral infection.

[0128] According to the detailed description of the invention, various other features or characteristics of the therapeutic target will be readily apparent in the embodiments.

[0129] 1. Antibodies and therapeutic products

[0130] 1.1 Test the following immune sera according to the method described below.

[0131] a. Human immune plasma was obtained from three individuals who received an influenza vaccine made from the 2009H1N1 influenza virus (pig) and three other individuals who were naturally infected with the 2009H1N1 influenza virus (pig) (China CDC National Influenza Center).

[0132] Antibody titers: 1:128-1:1280 (all adjusted to 1:128).

[0133] b. Human immune serum obtained from five individuals naturally infected with the 2009H1N1 influenza virus (pig) (China CDC National Influenza Center).

[0134] Antibody titer: 1:80-1:160 (all adjusted to 1:80).

[0135] c. Human immune serum obtained from five individuals who were naturally infected with seasonal H1N1 influenza virus (Guangdong / Baoan / 2006, Guangdong / Luohu / 2008, Tianjin / 2009) (National Influenza Center, Chinese Center for Disease Control and Prevention).

[0136] Antibody titers: 1:64-1:520 (adjusted to 1:128 if possible).

[0137] d. Human immune serum obtained from three individuals who were naturally infected with the H3N2 influenza virus (Brisbane / 59 / Xinjiang / 2007) (National Influenza Center, Chinese Center for Disease Control and Prevention).

[0138] Antibody titer: 1:1280 (all adjusted to 1:128).

[0139] e. Human immune serum obtained from three individuals naturally infected with H5N1 (avian) influenza virus (Anhui, 2005) (National Influenza Center, Chinese Center for Disease Control and Prevention).

[0140] Antibody titer: 1:64-1:152 (adjust to 1:128 if possible).

[0141] f. Human immune serum obtained from three individuals who were naturally infected with influenza B virus (Florida / 4 / Yunnan / 2007) (National Influenza Center, Chinese Center for Disease Control and Prevention).

[0142] g. A combination of human serum or plasma obtained from ten healthy individuals who were not infected with or vaccinated against influenza.

[0143] Antibody titer for seasonal H1N1 virus: 1:5.

[0144] 1.2 Test the following products according to the method described below.

[0145] a. A combination of human serum or plasma as described above.

[0146] b. Commercially available purified human immunoglobulin (Ig) (10%).

[0147] c. Preparations containing N-acetylneuraminic acid (1 mg / ml).

[0148] 1.3 Prepare the following serum mixture according to the method described below.

[0149] a. Mix 100 μL of human anti-09H1N1 (swine), anti-H1N1 (seasonal), and anti-H5N1 (avian) virus serum with 100 μL of human serum mixture.

[0150] b. Mix 100 μL of human anti-09H1N1 (swine), anti-H1N1 (seasonal), and anti-H5N1 (avian) virus serum with 100 μg of human Ig, respectively.

[0151] 100 μL of human anti-09H1N1 (swine), anti-H1N1 (seasonal), and anti-H5N1 (avian) virus serum were respectively mixed with 100 μL of a preparation containing N-acetylneuraminic acid as described above.

[0152] 2. Experimental models used to evaluate the safety of vaccines and antibodies.

[0153] 2.1 An animal model of chicken embryos and chicks

[0154] Chicken embryos at days 16 (E16) and 19 (E19) were treated with allantoic injection of various human anti-influenza virus immune sera or other anti-pathogen antibodies, and the health of the chicks was observed. The chicks hatched at approximately E21.

[0155] Embryos at day 16 (E16) and day 19 (E19) were treated by allantoic injection of 100 μL of the following: 1) saline alone (n=20); 2) human serum conjugate as described above (n=20); 3) human serum obtained from subjects infected with RSV (n=20); 4) human serum against 2009 H1N1 (swine) virus (n=40); 5) human serum against seasonal H1N1 virus (n=40); 6) human serum against H3N2 virus (n=40); 7) human serum against H5N1 (avian) virus (n=40); and 8) human serum against type B virus (n=40). Chicken embryos were incubated at 35°C until chicks hatched around day 21.

[0156] As described above, injections of sera against 2009H1N1 virus, seasonal H5N1 (avian) virus, and influenza B virus resulted in leg deformities in chicks, similar to human Guillain-Barre syndrome (GBS). The frequency of GBS-like symptoms induced by anti-influenza virus antibodies is listed in Table 1. These results indicate that antibodies induced by 2009H1N1 (swine) virus or vaccines containing 2009H1N1 (swine) virus have the highest risk of inducing GBS-like symptoms, followed by antibodies induced by avian influenza H5N1 (avian) virus and influenza B virus. However, with decreasing doses (50 μL or less), no serum induced significant symptoms in antibody-treated chickens. These data suggest that medium or high doses of antibodies against H1N1 (swine), H5N1, and influenza B viruses can induce serious adverse reactions in chicken embryos and chicks.

[0157] Table 1. Frequency of GBS-like symptoms in chicks injected with antibodies.

[0158]

[0159]

[0160] *: Serum pool of ten healthy individuals who were neither infected with nor vaccinated against influenza.

[0161] **: Serum obtained from subjects who were naturally infected or vaccinated against the 2009 H1N1 influenza virus.

[0162] RSV = Respiratory Syncytial Virus; GBS = Guillain-Barre Syndrome

[0163] In addition, injection of human anti-influenza virus serum resulted in fetal and neonatal death in chicken embryos or chicks. Mortality rates are listed in Table 2. Results using this animal model indicate that antibodies induced by influenza B virus have the highest risk of causing fetal and neonatal death (65.0%), followed by antibodies induced by avian influenza H5N1 virus (57.5%), 2009 H1N1 (swine) virus (52.5%), H3N2 virus (37.5%), and seasonal H1N1 virus (15.0%). As a control, anti-respiratory syncytial virus (RSV) serum did not cause significant GBS-like symptoms or death (Tables 1-2).

[0164] Table 2 Mortality frequency of chicks injected with antibodies

[0165]

[0166] *: Includes fetal and neonatal death

[0167] In another experiment, the serum mixture prepared in 1.3 above was also tested using an animal model. Injection of the serum mixture described above significantly reduced GBS-like symptoms or death in chicks compared to injection of a single dose of anti-influenza virus serum alone (Table 2). Preparations containing N-acetylneuraminic acid methyl ester as an N-acetylneuraminic acid analog were also tested using the method described above. The results were similar to those for N-acetylneuraminic acid as shown in Table 2.

[0168] The effective dose of N-acetylneuraminic acid or N-acetylneuraminic acid analogues (such as N-acetylneuraminic acid methyl ester) is 0.1 mg / kg to 100 mg / kg.

[0169] 2.2 Animal Model of Pregnant Rats

[0170] Pregnant mice were treated with intraperitoneal injection (IP) of multiple human anti-influenza virus immune sera or other anti-pathogen antibodies at gestational days E16 and E19, respectively. Newborn mice were born around E21, and their health status was observed.

[0171] As shown in Tables 3-4, injection of human anti-influenza virus serum into pregnant mice at days 16 (E16) and E19 of gestation resulted in abortion, premature birth, or prolonged labor in the pregnant mice, as well as fetal and neonatal mortality in newborn suckling mice born from the mother mice. The frequencies of fetal and neonatal mortality in newborn suckling mice are listed in Table 3. The results of this animal model indicate that antibodies induced by H5N1 (avian) virus have the highest risk of death (34.6%), followed by antibodies induced by seasonal H1N1 virus (26.7%), H3N2 virus (26.5%), influenza B virus (23.5%), and 2009 H1N1 (swine) virus (22.9%).

[0172] Table 3 Mortality rates in fetal or newborn mice injected with anti-influenza virus serum

[0173]

[0174] The abortion frequency in pregnant mice is listed in Table 4. The results of this animal model indicate that antibodies induced by influenza B virus have the highest risk of causing abortion (33.3%), followed by antibodies induced by 2009 H1N1 (swine) virus (31.3%), H3N2 virus (28.6%), seasonal H1N1 virus (21.4%), and H5N1 (avian influenza) virus (20.0%).

[0175] Table 4. Abortion frequency in pregnant mice injected with anti-influenza virus serum.

[0176]

[0177] Histological changes in dead mice were observed in the lungs of dead chickens and suckling mice. Figure 1 Erythrocyte infiltration, proliferation of inflammatory cells and epithelial cells, and localized tissue lesions were observed in the brain, heart, and liver. Additionally, human IgG binding to these organs was detected in tissue sections from dead chickens and suckling mice (data not provided).

[0178] In summary, this invention provides the first evidence that high levels of antibodies against influenza viruses are toxic to fetuses and newborns. Based on our common knowledge, this is the first direct evidence of a correlation between influenza virus-induced antibodies and serious adverse reactions such as GBS-like symptoms and fetal or neonatal death. This result suggests that influenza vaccination during pregnancy may pose risks to the fetus and newborn.

[0179] 3. Products used for the prevention and treatment of illnesses caused by harmful antibodies.

[0180] 3.1 Therapeutic products used for the prevention and treatment of fetal and neonatal death

[0181] The serum mixture prepared according to the method described in 1.3 above was injected intraperitoneally twice, at E16 and E19 days of gestation. Newborn mice were born at approximately E21 days, and their health status was observed.

[0182] Compared to injection of a single dose of anti-influenza virus serum alone, injection of the serum mixture described above into pregnant mice prevented fetal and neonatal mortality in these mothers (Table 5). Furthermore, no significant abortion, premature birth, or prolonged labor was observed in the mothers treated with the serum mixture. Preparations containing N-acetylneuraminic acid methyl ester as an N-acetylneuraminic acid analog were also determined using the N-acetylneuraminic acid method described above. The results obtained were similar to those for N-acetylneuraminic acid shown in Table 5.

[0183] Table 5. Therapeutic products for preventing fetal and neonatal death caused by influenza virus antibodies.

[0184]

[0185] Note: SP = serum conjugate; Ig = immunoglobulin; Neu5Ac = N-acetylneuraminic acid;

[0186] S = Seasonality; H = People.

[0187] Based on the obtained data and observations, the following products are effective in preventing or treating diseases or conditions caused by harmful antibodies (such as GBS-like conditions, fetal or neonatal death): 1) nonspecific serum from healthy individuals; 2) human immunoglobulins; and 3) N-acetylneuraminic acid or N-acetylneuraminic acid analogs (such as N-acetylneuraminic acid methyl ester).

[0188] For the prevention or treatment of diseases or conditions caused by harmful antibodies as described above (such as GBS-like conditions, fetal or neonatal death), the effective dose of serum or plasma is from about 0.1 ml / kg to 100 ml / kg; the effective dose of human Ig products is from about 0.1 mg / kg to 100 mg / kg; and the effective dose of N-acetylneuraminic acid or its analogues (such as N-acetylneuraminic acid methyl ester) is from 0.1 mg / kg to 100 mg / kg.

[0189] 3.2 Therapeutic products for treating influenza virus infection

[0190] Ten groups of newborn bulb / c mice on day 5 (P5) were inoculated with 30 μL (µL) of A / PR / 8 / 34 (H1N1) influenza virus (titer: 1:512, diluted 100-fold with physiological saline) via oral and nasal administration; and on day 6 (P6), 100 µL of physiological saline containing 1) saline alone; 2) 100 µL of human serum conjugate; 3) 100 µL of human Ig; and 4) 30 µL of human anti-2009H1N1 (swine) was administered intraperitoneally. Viral serum; 5) 30 μL human anti-H1N1 serum; 6) 30 μL human anti-H5N1 serum; 7) 30 μL human anti-2009H1N1 (swine) virus serum plus 100 μL N-acetylneuraminic acid (Neu5Ac); 8) 30 μL human anti-H5N1 serum plus 100 μL Neu5Ac; 9) 100 μL human serum mixture plus 100 μL Neu5Ac; and 10) 100 μL human Ig plus 100 μL Neu5Ac. Mice were fed for 7 days post-infection.

[0191] Table 6. Treatment products for influenza virus infection caused by A / PR / 8 / 34 (H1N1) virus.

[0192]

[0193] Note: Ig = Immunoglobulin; Neu5Ac = N-acetylneuraminic acid; S = Seasonality.

[0194] According to the summary in Table 6, 17 / 20 (87%) of suckling mice treated with saline, and all suckling mice treated alone with anti-2009H1N1 serum or anti-seasonal H1N1 serum, died on day 3 post-infection. 50% or more of the suckling mice treated with human serum conjugates (50%) or human Ig or anti-H5N1 serum survived. The mortality rate was significantly reduced in suckling mice treated with serum conjugates containing N-acetylneuraminic acid (Table 6). This result suggests that N-acetylneuraminic acid can significantly reduce the toxicity of harmful antibodies or increase the efficacy of human Ig or human serum or plasma in treating severe influenza virus infections. The data also indicate that high doses of antibodies against 2009H1N1 (swine) virus alone are harmful, not beneficial, in the treatment of influenza virus infections.

[0195] For the treatment of influenza virus infection, the effective dose of serum or plasma is from about 0.1 ml / kg to 100 ml / kg; the effective dose of human Ig products is from about 0.1 mg / kg to 100 mg / kg; and the effective dose of N-acetylneuraminic acid or N-acetylneuraminic acid analogs (such as N-acetylneuraminic acid methyl ester) is from 0.1 mg / kg to 100 mg / kg.

[0196] 3.3 Treatment products for the prevention of influenza virus infection

[0197] In another experiment, using the mouse model described in 3.2, ten groups of newborn 5-day-old (P5) bulb / c suckling mice were pretreated by intraperitoneal injection of serum and serum mixture as described in 3.2 above. The suckling mice were orally inoculated with 30 μl of A / PR / 8 / 34 (H1N1) virus (titer: 1:512, diluted 100-fold with physiological saline) on day 6 (P6) and were fed for 7 days post-viral infection.

[0198] The results are summarized in Table 7. The data indicate that 1) human serum compositions or human immunoglobulin or anti-H5N1 alone reduced the mortality rate from A / PR / 8 / 34 (H1N1) infection from 87.0% to 50% or 42.9% (1.74-2.0 times); 2) specific serum against seasonal H1N1 or specific serum against 2009H1N1 (swine) virus (containing specific antibodies at a titer of 1:128) was ineffective or harmful in preventing A / PR / 8 / 34 (H1N1) infection; 3) the combined use of N-acetylneuraminic acid with nonspecific human serum, or nonspecific human immunoglobulin, or specific anti-influenza virus antibodies, and N-acetylneuraminic acid was significantly effective in preventing H1N1 influenza virus infection; and 4) N-acetylneuraminic acid significantly improved the anti-infective efficacy of nonspecific human serum, or nonspecific human immunoglobulin, or specific anti-influenza virus antibodies, and reduced the toxicity of specific harmful antibodies (such as anti-influenza virus antibodies).

[0199] Table 7. Treatment products for the prevention of influenza virus infection with A / PR / 8 / 34 (H1N1) virus.

[0200]

[0201] Note: Ig = Immunoglobulin; Neu5Ac = N-acetylneuraminic acid; S = Seasonality.

[0202] For the prevention of influenza virus infection, the effective dose of serum or plasma is from about 0.1 ml / kg to 100 ml / kg; the effective dose of human Ig products is from about 0.1 mg / kg to 100 mg / kg; and the effective dose of N-acetylneuraminic acid or N-acetylneuraminic acid analogs (such as N-acetylneuraminic acid methyl ester) is from 0.1 mg / kg to 100 mg / kg.

[0203] 3.4 Another experimental model

[0204] A mouse model was established by injecting antibodies against the primary pathogen into animals and then infecting them with a secondary infectious pathogen.

[0205] Six groups of newborn 5-day-old (P5) bulb / c suckling mice were pretreated with antibodies against other pathogens via intraperitoneal injection. These antibodies did not react with the influenza virus described below. On day 6 (P6), the suckling mice were orally inoculated with 30 μl (µL) of A / PR / 8 / 34 (H1N1) virus (titer: 1:512, diluted 100-fold with physiological saline) and kept for 7 days post-infection. The antibodies against other pathogens tested were:

[0206] a. Human immune serum obtained from subjects infected with RSV;

[0207] b. Human immune serum obtained from subjects infected with HAV;

[0208] c. Human immune serum obtained from subjects infected with HBV;

[0209] c. Human immune serum obtained from subjects infected with rotavirus;

[0210] None of the antibodies reacted with the influenza virus.

[0211] The results are summarized in Table 8. Compared with suckling mice pretreated with saline, suckling mice pretreated with antibodies against HAV, HBV, RSV, and rotavirus were either not infected with influenza virus or had mild influenza virus infection. This data suggests that antibodies against the primary pathogen (such as HAV) can prevent infection by the secondary pathogen.

[0212] In addition, the combined use of N-acetylneuraminic acid with specific antibodies against pathogens different from influenza viruses significantly improved the efficacy of antibodies used to prevent H1N1 influenza virus infection.

[0213] Table 8. Multiple antibodies used to prevent influenza virus infection with A / PR / 8 / 34 (H1N1) virus.

[0214]

[0215] Note: Neu5Ac = N-acetylneuraminic acid; S = seasonal.

[0216] For the prevention of influenza virus infection, the effective dose of specific antibodies against pathogens other than influenza virus is from about 0.001 mg / kg to 100 mg / kg; the effective dose of N-acetylneuraminic acid or N-acetylneuraminic acid analogues (such as N-acetylneuraminic acid methyl ester) is from 0.1 mg / kg to 100 mg / kg.

[0217] 3.4 Combined use of therapeutic antibodies and N-acetylneuraminic acid

[0218] As described above and shown in Tables 6-8, the combined use of N-acetylneuraminic acid with nonspecific human serum, or nonspecific human immunoglobulin, or specific antibodies against influenza viruses, or specific antibodies against non-influenza viruses, has a significant effect on preventing H1N1 influenza virus infection. This data indicates that N-acetylneuraminic acid significantly enhances the anti-infective efficacy of nonspecific human serum, or nonspecific human immunoglobulin, or specific antibodies against influenza viruses or specific antibodies against pathogens different from influenza viruses, and reduces the toxicity of specific harmful antibodies (such as anti-influenza virus antibodies).

[0219] Additional Definitions

[0220] As used herein, the term "treating" or "treatment" refers to a clinical intervention (e.g., administration of immunoglobulin products, serum, or plasma as described herein) aimed at altering the natural course of a treated individual or cell in the clinicopathological process of a viral infection. The intended effects of treatment include reducing the rate of disease progression or mortality, improving or alleviating the disease state, and alleviating or improving prognosis. In some embodiments, treatment may improve symptoms of a viral infection (such as influenza virus infection), reduce the frequency or severity of illness caused by a viral infection (such as influenza virus), and / or improve symptoms exhibited by the patient (e.g., flu symptoms, including, but not limited to, fever, chills, cough, sore throat, body aches, and fatigue). A response is achieved when the patient experiences partial or complete relief or a reduction in signals or symptoms of the disease, and, in some embodiments, includes survival. For example, a subject is successfully "treated" if one or more symptoms associated with a viral infection (such as influenza virus) are relieved or eliminated.

[0221] As used herein, the terms “preventing” or “prevention” include providing protection against the occurrence or recurrence of a viral infection (such as influenza virus) within an individual. An individual may be predisposed to infection or susceptible to viral infection (such as through influenza virus infection) but has not yet been infected with the virus.

[0222] As used in this article, an individual at “risk” for viral infection (such as influenza virus infection) refers to an individual who is likely to be exposed to a viral pathogen, or an individual who, if infected with a viral pathogen, has one or more risk factors for a severe response.

[0223] "Effective dose" refers to the lowest dose, at the necessary dosage and time, of an immunoglobulin product, serum, or plasma as described herein to achieve the intended or prescribed effect, including therapeutic or preventative outcomes. The effective dose may be provided in one or more administrations.

[0224] As used herein, the terms "patient" or "individual" refer to a mammal, including, but not limited to, humans or non-human mammals such as cattle, horses, pigs, dogs, sheep, or cats. In some embodiments, the patient or individual is a human. In some embodiments, the patient does not require a blood transfusion because, for example, due to injury or a bleeding clotting disorder, such as, but not limited to, hemophilia, von Willebrand disease, and leukemia. In some embodiments, both the patient and the healthy individual are humans. In some embodiments, both the patient and the healthy individual are cattle, sheep, chickens, pigs, horses, dogs, or cats.

[0225] As used herein, patients in this invention include male or female human or non-human mammals who have or are at risk of developing diseases and conditions caused by harmful antibodies, newborns, infants aged 1-12 months, adolescents aged 1-18 years, adults, pregnant and lactating females, and pregnant and lactating females with a fetus or lactating infant.

[0226] Additional Implementation Methods

[0227] Another embodiment of the present invention discloses a therapeutic product containing a vaccine or an inactivated pathogen for treating severe infections caused by antibodies induced during infection; wherein the therapeutic vaccine or inactivated pathogen can neutralize harmful antibodies induced during infection.

[0228] Another embodiment of the present invention discloses a method of administering a therapeutic vaccine or an inactivated pathogen to a patient to prevent or treat a disease or condition caused by antibodies against the vaccine or pathogen; wherein the antibodies are induced during infection; and the therapeutic vaccine or inactivated pathogen can neutralize the harmful antibodies induced during infection.

[0229] Other embodiments besides those described above will be readily apparent. Therefore, these terms and expressions are used only to describe what has been disclosed through embodiments, and do not limit the scope of the disclosure. It is anticipated that other embodiments different from the foregoing and not departing from the spirit and scope of the description and claims herein will be readily apparent. All patents, published patents, and other references cited herein are incorporated herein by reference in their entirety.

Claims

1. The use of N-acetylneuraminic acid in the preparation of medicaments for treating or preventing viral pathogen infection, wherein the active ingredient of the medicament also includes nonspecific serum at an effective dose for a patient, the patient being a patient already diagnosed with or at risk of viral pathogen infection, wherein... The nonspecific serum was obtained from a healthy individual; The viral pathogen is influenza virus H1N1; the ratio of nonspecific serum to N-acetylneuraminic acid at a concentration of 1 mg / ml is 100 μL: 100 μL.

2. The application according to claim 1, wherein, The nonspecific serum may be in the form of a solution, lyophilized form, injectable form, infusion form, or in the form coupled to nanoparticles.

3. The application according to claim 1, wherein, The concentration of N-acetylneuraminic acid in the drug is between 0.01 and 100 mg / g.

4. The application according to claim 1, wherein, The concentration of N-acetylneuraminic acid in the drug is between 0.01 and 100 mg / ml.

5. The application according to claim 1, wherein, The nonspecific serum is administered via subcutaneous injection, topical application, oral administration, intramuscular injection, intravenous injection, intracavitary injection, or inhalation.

6. The application according to claim 5, wherein, The intracavitary injection is intraperitoneal injection; or the topical administration is transdermal administration.

7. The application according to claim 1, wherein, The patients and healthy individuals in question are all of the same species of mammal.

8. The application according to claim 6, wherein, Both the patients and healthy individuals mentioned are human beings.

9. The application according to claim 1 or 8, wherein, The effective dose of the nonspecific serum is from 0.1 ml / kg to 100 ml / kg; And / or the effective dose of the N-acetylneuraminic acid is from 0.1 mg / kg to 100 mg / kg.

10. A product for treating and preventing infections caused by viral pathogens or diseases or symptoms caused by harmful antibodies; wherein, The product is in the form of a composition, and the active ingredient of the composition is: N-acetylneuraminic acid, and Nonspecific serum; The harmful antibodies are either induced during infection or vaccination, or are passively introduced therapeutic antibodies. Diseases or conditions caused by harmful antibodies include infectious diseases, serious adverse reactions to vaccines or therapeutic antibodies, infection-related autoimmune diseases, allergies and inflammation, and infection-related tumors. The nonspecific serum is obtained from healthy individuals. The pathogen of the virus is influenza virus H1N1; The ratio of nonspecific serum to N-acetylneuraminic acid at a concentration of 1 mg / ml is 100 μL: 100 μL.

11. The product according to claim 10, wherein, Serious adverse reactions to infection and vaccination include Guillain-Barré syndrome-like symptoms, death, miscarriage, premature or prolonged birth, and neonatal death.

12. The product according to claim 10, wherein, The infection is an influenza virus infection; the vaccination is an influenza virus vaccination.

13. The product according to claim 10, wherein, The product is in the form of a composition containing 0.1-1000 ml of serum.

14. The product according to claim 10, wherein, The product is in the form of a composition containing about 0.1-20 mg / ml or about 0.1-20 mg / g of N-acetylneuraminic acid and 0.1-1000 ml of serum.

15. The product according to claim 10, wherein, The product is in the form of a solution, a lyophilized form, an injection form, an infusion form, or a form coupled to nanoparticles.

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