双价痢疾菌苗滴鼻免疫小鼠诱导粘膜与系统免疫反应

目的探讨双价痢疾菌苗滴鼻免疫后对小鼠不同粘膜部位和系统免疫部位的影响.方法BALB/c小鼠随机分为3组,每组20只,FSM-2117或FS-54164×107CFU/只经滴鼻途径免疫小鼠,间隔2w,3次免疫后7d活杀,分离NALT、鼻通道、脾、小肠PP及MLN淋巴细胞,采用流式细胞术检测其淋巴细胞表型的变化;收集鼻咽、肺、肠、生殖道冲洗液和血清,采用ELISA法检测其中特异性抗福氏、宋内氏LPSIgA和IgG.结果两株菌苗经鼻内免疫都诱发了鼻咽、肺、胃肠道和生殖道等不同粘膜部位及血清中特异性抗福氏、宋内氏LPSIgA、IgG的显著增加(P＜0.01);NALT、NP和PP淋巴细胞中CD3+T细胞显著增加,其中以CD4+T细胞增加为主;FSM-2117免疫组的脾细胞中B220+细胞显著增加,而FS-5416免疫组的脾细胞中CD3+T细胞显著增加.结论两株菌苗经鼻粘膜免疫均可诱导不同粘膜部位及系统免疫反应的发生,鼻粘膜是一个安全有效的免疫途径.     

Aujeszky's disease in the guinea pig: Cellular and humoral responses following immunization

Summary The fatal disease caused by virulent ADV in guinea pigs was found to be identical to that seen in sheep and cattle. Intramuscular (i.m.) injection of an avirulent strain of ADV (Bartha) yielded better immunity to challenge after 3 weeks than did intranasal (i.n.) immunization, and this was reflected in differences in histopathological changes in the brain.Serum antibodies active in antibody-dependent cell-mediated cytotoxicity (ADCC) were titrated using polymorphonuclear leukocytes as effector cells. ADCC correlated fairly well with virus neutralization and was a far more sensitive technique. There was good, but not complete, correlation between ADCC and protection. Lymphocyte responsiveness to virus antigensin vitro was assessed by³H-thymidine uptake and lymphokine tests. Lymphocyte stimulation and mitogenic factor responses were low grade but blood lymphocyte stimulation was more pronounced in the better-protected animals. Macrophage migration inhibition correlated neither with serum ADCC nor with protection, being equally demonstrable in the two immunized groups.This work was carried out while this establishment under its former title Microbiological Research Establishment was under the management of the Ministry of Defence.     

Local and systemic immune responses following oral immunization of foetal lambs.

Foetal lambs were immunized orally 6-15 days before birth by introducing horse spleen ferritin into the amniotic fluid. Immunized and non-immunized lambs were killed at birth, usually before they had suckled, blood and intestinal contents were collected and single cell suspensions were prepared from spleen, mesenteric lymph nodes and jejunum. Specific antibody was detected in serum and intestinal contents of all immunized lambs which had not suckled. Specific antibody was usually not detected in samples from non-immunized lambs. In immunized lambs antibody activity in serum was associated with IgM and in intestinal contents with IgA and IgM. In agreement with these findings, the levels of IgM and IgA in serum and intestinal contents of immunized lambs were relatively high. Generally, immunoglobulins were not detected in samples from non-immunized lambs. Relatively high proportions of cells secreting specific antibody were present in the tissues of immunized but not non-immunized lambs. In the spleen most of the cells were secreting IgM antibody, in mesenteris lymph nodes IgM cells predominated and small numbers of IgA cells were detected, and in the jejunum approximately equal numbers of IgA and IgM cells were secreting specific antibody.     

Protection against keratoconjunctivitis shigellosa induced by immunization with outer membrane proteins of Shigella spp.

Active immunization of guinea pigs and rabbits with outer membrane proteins (OMP) isolated from Shigella flexneri 3a and Shigella sonnei phase I protected the animals against keratoconjunctivitis shigellosa induced with the homologous or heterologous strain. Protection was also achieved in rabbits after passive immunization with anti-OMP immune serum. Active immunization with lipopolysaccharide of S. flexneri 3a did not protect rabbits against keratoconjunctivitis shigellosa.     

Primary immune response of guinea pig spleen cells in vitro

Spleen cells of the primed guinea pig, when cultured alone, failed to make a primary immune response against heterologous erythrocytes. When supplemented with lymphocytes obtained from mineral oil induced peritoneal exudate (PEL), a primary immune response against sheep, horse and chicken RBC was obtained. The kinetics, dose response and requirement of PELs for this response are reported.     

T-lymphocyte response in a guinea pig model of tuberculous pleuritis.

The ability to induce tuberculous pleuritis in Mycobacterium bovis BCG-vaccinated guinea pigs was investigated as a model of human disease. A pleural effusion of 5 to 10 ml was obtained 6 to 7 days after the bilateral pleural injection of a suspension of heat-killed M. tuberculosis cells. Histological lesions were indicative of granulomatous pleuritis. Comparative studies of T lymphocytes obtained from pleural fluid and peripheral blood revealed increased antigen-driven lymphoproliferation and E rosette formation in pleural effusion lymphocytes. The CD2+ T-lymphocyte population appeared to be expanded or concentrated in pleural fluid, suggesting a compartmentalization of antigen-reactive T lymphocytes. These data demonstrate that experimental tuberculous pleuritis with effusion, closely resembling the human disease, can be produced in BCG-vaccinated guinea pigs.     

Salmonella infections: immune and non-immune protection with vaccines.

Salmonella enterica in poultry remains a major political issue. S. enterica serovar Enteritidis, particularly, remains a world-wide problem. Control in poultry by immunity, whether acquired or innate, is a possible means of containing the problem. Widespread usage of antibiotics has led to the emergence of multiple antibiotic-resistant bacteria. This problem has indicated an increasing requirement for effective vaccines to control this important zoonotic infection. An attempt is made in the present review to explain the relatively poor success in immunizing food animals against these non-host-specific Salmonella serotypes that usually produce food-poisoning, compared with the success obtained with the small number of serotypes that more typically produce systemic "typhoid-like" diseases. New examinations of old problems such as the carrier state and vertical transmission, observed with S. Pullorum, is generating new information of relevance to immunity. Newer methods of attenuation are being developed. Live vaccines, if administered orally, demonstrate non-specific and rapid protection against infection that is of biological and practical interest. However, from the point of view of consumer safety, there is a school of thought that considers inactivated or sub-unit vaccines to be the safest. The benefits of developing effective killed or sub-unit vaccines over the use of live vaccines are enormous. Recently, there have been significant advances in the development of adjuvants (e.g. microspheres) that are capable of potent immuno-stimulation, targeting different arms of the immune system. The exploitation of such technology in conjunction with the ongoing developments in identifying key Salmonella virulence determinants should form the next generation of Salmonella sub-unit vaccines for the control of this important group of pathogens. There are additional areas of concern associated with the use of live vaccines, particularly if these are generated by genetic manipulation.     

Intransal or intragastric immunization with proteosome-Shigella lipopolysaccharide vaccines protects against lethal pneumonia in a murine model of Shigella infection.

Mice immunized intransally or intragastrically with proteosome vaccines containing either Shigella sonnei or S. flexneri 2a lipopolysaccharide were protected against lethal pneumonia caused by homologous organisms in an experimental murine intranasal challenge model of Shigella infection. Histopathological analysis demonstrated that immunization also protected against the progressive lesions resulting from invasion of the pulmonary mucosa by S. sonnei. These data show that mucosal proteosome-lipopolysaccharide vaccines can protect against lethal bacterial pneumonia and indicate that such vaccines are promising candidates for protection against intestinal shigellosis.     

Long-term protection after immunization with protein-polysaccharide conjugate vaccines in infancy

The polysaccharide-encapsulated bacteria, Haemophilus influenzae type b, Neisseria meningitidis and Streptococcus pneumoniae are important causes of invasive bacterial infection in childhood, accounting for most of the cases of bacterial pneumonia and meningitis worldwide. Protein-polysaccharide conjugate vaccines have been developed over the last 20 years and have proven very effective in controlling these infections. Although studies have consistently shown that herd immunity is critical for population protection, long-term individual protection against polysaccharide-encapsulated bacteria appears to depend on persisting antibody and, perhaps to a lesser extent, immunological memory. However, some studies have reported that the concentration of serum antibody and vaccine effectiveness are not sustained after infant immunization, despite persistence of immunological memory. In this article, we detail the mechanisms of protection against invasion by encapsulated bacteria, describe the age-dependent B-cell and antibody responses to protein-polysaccharide conjugate vaccines and propose strategies to guarantee protection during periods of increased disease burden.     

Outbreak of keratoconjunctivitis due to Salmonella weltevreden in a guinea pig colony.

The purpose of this report is to demonstrate that the ability to produce keratoconjunctivitis (KC) is a property found in Salmonella weltevreden. This observation is contrary to previous reports that Salmonella spp. do not produce KC. An outbreak of KC due to S. weltevreden occurred in a guinea pig colony, and the animals carried the organism in the intestinal tract. The same Salmonella serotype that caused an epidemic of diarrhea in humans and a routine laboratory isolate also possessed the ability to induce KC. Unlike Shigella spp. (the prototype organisms positive for KC), S. weltevreden induced KC and bound Congo red dye even when grown at 30 degrees C. It invaded HeLa cells in culture but did not hybridize with a DNA probe for invasiveness of Shigella spp. and enteroinvasive Escherichia coli even though it harbored plasmids. It was susceptible to all the antibiotics tested, was hydrophobic, and showed mannose-sensitive hemagglutination. It did not have enterotoxic or cytotoxic activities.     

Specific immune response in the human respiratory tract following oral immunization with live typhoid vaccine

Specific antibody responses in the lower respiratory tract of human subjects to orally administered Salmonella typhi Ty21a are reported. These responses, predominantly of the immunoglobulin G class, were determined to be a transudate from serum. These results were supported by the similarity in responses to parenteral administration of heat-killed typhoid vaccine. Specific immunoglobulin A antibody was a poor contributor to the respiratory antibody response to either vaccine.     

Serum immune response to Shigella protein antigens in rhesus monkeys and humans infected with Shigella spp.

The serum antibody response to proteins encoded by the virulence-associated plasmid of Shigella flexneri was determined in monkeys challenged with virulent S. flexneri serotype 2a. With water-extractable antigen in an enzyme-linked immunosorbent assay, a significant increase in antibody titer against proteins from a plasmid-carrying, virulent strain of S. flexneri serotype 5 could be demonstrated in convalescent sera. There were minimal antibody titers against proteins from an avirulent (plasmid-free) organism. Previously identified plasmid-coded polypeptides a, b, c, and d were predominant antigens recognized by a majority of the convalescent sera in immunoblots. An additional 140-megadalton plasmid-coded polypeptide was also recognized by half of the sera. Convalescent serum from an infected monkey recognized antigens on the bacterial surface in several different plasmid-containing Shigella species and in an enteroinvasive Escherichia coli strain. A survey of sera obtained from children 5 to 10 years of age who had been infected with S. flexneri or S. sonnei revealed high enzyme-linked immunosorbent assay titers in both acute and convalescent sera against a water extract from a virulent Shigella strain. In contrast, children under 3 years of age had no antibody titer in either acute or convalescent sera against the virulence-associated shigella proteins, while 3- to 4-year-old children mounted an immune response against these proteins only in convalescence.     

Delayed hypersensitivity to myxoviruses and myxovirus vaccines in the guinea pig

Influenza, mumps and measles viruses were examined for their ability to induce in guinea pigs homologous and cross-reactive delayed hypersensitivity. A majority of the animals skin tested with homologous and a portion of the animals skin tested with heterologous viruses and vaccines developed positive reactions. Findings with the heterologous preparations suggest that the observed cross-reactive hypersensitivity might be due to shared antigens of viral or substrate origin in the influenza, mumps and measles preparations. The present findings in guinea pigs suggest that the adverse effects, attributed in whole or in part to induced hypersensitivity, observed in man following injection of killed influenza, mumps and measles vaccines could be due to cross-reactive delayed hypersensitivity resulting from the use of these preparations.     

痢疾菌苗滴鼻免疫小鼠诱导不同部位淋巴组织的免疫应答

目的 探讨痢疾菌苗滴鼻免疫后，小鼠不同粘膜部位和其它部位免疫应答的影响。方法将Ｂａｌｂ／ｃ小鼠随机分为３组，每组１５只。舰疾菌苗株 ＴＩＭ－２１１７或 ＦＳ－５４１６，按 ５×10～6、１×１０～７、４×１０～７和４×１０～７ＣＦＵ／只滴鼻跟 Ｂａｌｂ／ｃ小鼠４次涧隔两 ｗｂ。分离鼻相关淋巴组织（ＮＡＬＴ）、鼻通道（ＮＰ）及脾、小肠的ＰＰ结淋巴细胞。一部分细胞采用流式细胞术检测淋巴细胞表型的变化；另一部分细胞与全菌破碎抗原共培养，于不同时间点取出，提取ＲＮＡ做ＲＴ－ＰＣＲ。结果两株菌苗经鼻内免疫后，ＮＡＬ、ＮＰ和ＰＰ结的淋巴细胞中ＣＤ３Ｔ细胞显著增加，其中以ＣＤ４＋Ｔ细胞增加为主。ＴＳＭ－２１１７株免疫组的脾细胞中，Ｂ２２０＋细胞显著增加；而ＦＳ－５４１６株免疫组的脾细胞中，ＣＤ３＋Ｔ细胞显著增加。免疫小鼠的ＮＡＬＴ、ＮＰ和ＰＰ结淋巴细胞，在体外经抗原刺激后，均在不同时间出现ＩＦＮ－γ和ＩＬ-４ｍＲＮＡ的表达，ＴＧＦ－β ｍＲＮＡ的表达在免疫前后无明显变化。结论痢疾菌苗经鼻粘膜免疫，可诱导不同粘膜部位及全身免疫应答的产生。     

Contraction of guinea pig trachea with antibodies to guinea pig IgE. An in vitro model for asthma

Rabbit antiserum to guinea pig IgE was prepared. This antiserum absorbed IgE antibodies to dinitrophenyl determinants when examined by passive cutaneous anaphylaxis. This antiserum also provoked contraction of tracheal rings from normal guinea pigs in vitro. This system is a new model for asthma in which only IgE among immunoglobulins reacts.     

Development of a guinea pig immune response-related microarray and its use to define the host response following Mycobacterium bovis BCG vaccination

Immune responses in the guinea pig model are understudied because of a lack of commercial reagents. We have developed a custom-made guinea pig oligonucleotide microarray (81 spots) and have examined the gene expression profile of splenocytes restimulated in vitro from Mycobacterium bovis BCG-vaccinated and naive animals. Eleven genes were significantly (P < 0.05) up-regulated following vaccination, indicating a Th1-type response. These results show that microarrays can be used to more fully define immune profiles of guinea pigs.     

Hemagglutinin and neuraminidase specific cell-mediated immune responses to influenza A virus following immunization in guinea pigs

Groups of guinea pigs were immunized with different inactivated recombinant influenza A viruses including H3ChN2Ch, Heq1N2Ch, H3ChNeq1 or uninfected allantoic fluid. Employing hemagglutination and neuraminidase inhibition tests, an in-vitro lymphocyte transformation (LTF) assay, and rosetting techniques for the separation of lymphocytes, influenza hemagglutinin (HA) and neuraminidase (NA) specific antibody and cell-mediated immune (CMI) responses were evaluated. Inactivated H3ChN2Ch, H3ChNeq1, HavN2Ch, and Heq1Neq1 recombinant influenza viruses were used as test antigens. Following immunization the CMI and antibody responses to influenza were characterized by the induction of specific LTF and antibody activity to homotypic HA or NA antigens but not to heterotypic HA or NA antigens. The temporal kinetics of the antibody response to influenza antigens was characterized by a prompt onset being initially detected at 1-2 weeks and reaching peak titers 3-4 weeks after immunization. Influenza specific LTF responses were first detected one week after immunization and declined to minimal responses at eight weeks. T-lymphocytes but not B-lymphocytes were capable of in-vitro recognition of the HA and NA antigens. After recognition the subsequent in-vitro lymphoproliferation was shown to involve both T and B lymphocytes.