Influences of anti-mouse interleukin-6 receptor antibody on immune responses in mice

In the present study, we examined the effect of anti-IL-6 receptor antibody (MR16-1) on humoral and cellular immune responses in mice. MR16-1 did not affect antigen-specific antibody production in either the primary or secondary response in mice immunized with dinitro-phenyl (DNP)-keyhole limpet haemocyanin (KLH) in saline. DNP-KLH immunization with complete Freund's adjuvant (CFA) markedly augmented anti-DNP antibody production and induced interleukin 6 (IL-6) production in serum. In this case, MR16-1 significantly suppressed antibody production and further increased serum IL-6 levels. Regarding the cellular response, we studied the effect on the delayed-type hypersensitivity (DTH) response. DTH response was induced in mice by the immunization with Mycobacterium butyricum with incomplete Freund's adjuvant and following antigen challenge into the footpad 14 days after immunization. When MR16-1 was injected immediately after immunization, the DTH response was significantly suppressed and enlargement of the spleen was also suppressed. This suppressive effect was observed, when MR16-1 was administered on day 0, but not on days 5 and 10. Again, serum IL-6 levels were much higher in MR16-1-treated mice compared with controls. Furthermore, spleen cells from control mice released IL-2 and INFgamma by the stimulation of antigen in vitro. In contrast, spleen cells from MR16-1-treated mice produced these cytokines at a marginal level. In contrast, MR16-1 did not suppress the DTH response, when it was injected immediately after antigen challenge. Our results suggest that IL-6 does not always involve antibody production, although IL-6 augments antibody production, and that IL-6 is essential for the induction of Th1 cells.     

Human respiratory syncytial virus: pathogenesis,immune responses,and current vaccine approaches

Respiratory syncytial virus continues to pose a serious threat to the pediatric populations worldwide. With a genomic makeup of 15,200 nucleotides, the virus encodes for 11 proteins serving as envelope spikes, inner envelope proteins, and non-structural and ribonucleocapsid complexes. The fusion (F) and attachment (G) surface glycoproteins are the key targets for neutralizing antibodies. The highly variable G with altered glycosylations and the conformational alternations of F create challenges for vaccine development. The metastable F protein is responsible for RSV-host cell fusion and thus infectivity. Novel antigenic sites were identified on this form following its stabilization and solving its crystal structure. Importantly, site ø displays neutralizing activity exceeding those of post-F-specific and shared antigenic sites, such as site II which is the target for Palivizumab therapeutic antibody. Induction of high neutralizing antibody responses by pre-F immunization in animal models promoted it as a major vaccine candidate. Since RSV infection is more serious at age extremities and in individuals with undermining health conditions, vaccines are being developed to target these populations. Infants below three months of age have a suppressive immune system, making vaccines’ immunogenicity weak. Therefore, a suggested strategy to protect newborns from RSV infection would be through passive immunity of maternal antibodies. Hence, pregnant women at their third trimester have been selected as an ideal target for vaccination with RSV pre-F vaccine. This review summarizes the different modes of RSV pathogenesis and host’s immune response to the infection, and illustrates on the latest updates of vaccine development and vaccination approaches.     

The common mucosal immune system and current strategies for induction of immune responses in external secretions

The selective induction of antibodies in external secretions is desirable for the prevention of various systemic as well as predominantly mucosa-restricted infections. An enormous surface area of mucosal membranes is protected primarily by antibodies that belong, in many species, to the IgA isotype. Such antibodies are produced locally by large numbers of IgA-containing plasma cells distributed in subepithelial spaces of mucosal membranes and in the stroma of secretory glands. In humans and in some animal species, plasma-derived IgA antibodies do not enter external secretions in significant quantities and systemically administered preformed IgA antibodies would be of little use for passive immunization. Systemic administration of microbial antigens may boost an effective S-IgA immune response only in a situation whereby an immunized individual had previously encountered the same antigen by the mucosal route. Local injection of antigen in the vicinity of secretory glands is usually accompanied by an undesirable concomitant systemic response and frequently requires the addition of adjuvants that are unacceptable for administration in humans. Immunization routes that involve ingestion or possibly inhalation of antigens lead to the induction of not only local but also generalized immune responses manifested by the parallel appearance of S-Iga antibodies to ingested or inhaled antigens in secretions of glands distant from the site of immunization. Based on extensive studies in animal models as well as in humans, convincing evidence is available that antigen-sensitized and IgA-committed precursors of plasma cells from GALT are disseminated to the gut, other mucosa-associated tissues, and exocrine glands. However, due to the limited absorption of desired antigens from the gut lumen of orally immunized individuals, repeated large doses of antigens are required for an effective S-IgA response. Novel antigen delivery systems for the stimulation of such responses are currently being examined in several laboratories. Live attenuated or genetically manipulated bacteria expressing other microbial antigens have also been used for selective colonization of gut-associated lymphoid tissues. Unique antigen packaging and the use of adjuvants suitable for oral administration hold promise for an efficient antigen delivery to critical tissues in the intestine and deserve extensive exploration. The oral immunization route appears to have many advantages over systemic immunization.(ABSTRACT TRUNCATED AT 400 WORDS)     

The human immune system in hu-PBL-SCID mice

Severe combined immunodeficiency (SCID) mice can be stably grafted with human peripheral blood lymphocytes, creating hu-PBL-SCID chimeras; essentially, these are mice with a human immune system. Here, Magdalena Tary-Lehmann, Andrew Saxon and Paul Lehmann discuss the immunobiology of these chimeras. The authors propose that hu-PBL-SCID chimerism evolves in two phases. During the first three weeks after grafting, many of the injected cells survive and the human immune system is functional. Subsequently, anti-mouse-reactive clones are selected and the immune system becomes nonfunctional. The implications of this scenario for the utilization of the hu-PBL-SCID model are discussed.     

SCID小鼠体内人体肿瘤移植及其人体免疫功能重建

目的观察高转移性人肺巨细胞癌系（ＰＧ）及转染了白细胞介素６基因的ＰＧ细胞（ＰＧＴＳ７）在ＳＣＩＤ小鼠及免疫重建ＳＣＩＤ小鼠体内的生长及转移行为,阐明ＰＧＴＳ７自分泌的白细胞介素６增强人体免疫的抗肿瘤作用。方法皮下移植人体肿瘤,部分受体鼠同时经腹腔注射人体外周血淋巴细胞。观察成瘤潜伏期、成瘤率、生长速度、体积、肺及淋巴结转移率和小鼠血清中人体免疫球蛋白（ＨＩｇ）的含量。结果全部动物见肿瘤生长,但ＰＧＴＳ７在免疫重建鼠体内成瘤潜伏期延长,肿瘤体积小、淋巴结转移率低,而血清中ＨＩｇ含量高。结论（１）ＳＣＩＤ小鼠亦为研究ＰＧ及ＰＧＴＳ７生长及转移的良好动物。（２）ＰＧＴＳ７自分泌白细胞介素６能刺激小鼠体内人淋巴细胞增殖、活化,增加ＨＩｇ的产生,降低肿瘤生长及淋巴结转移。     

Human immune responses to oral microorganisms: patterns of systemic antibody levels to Bacteroides species.

Human systemic antibody levels to oral members of the Bacteroides genus were assessed with an enzyme-linked immunosorbent assay. Antibody levels to B. gingivalis, two homology groups of B. intermedius, B. melaninogenicus, B. denticola, B. loescheii, B. corporis, B. oralis, B. buccae, and B. gracilis were determined in subjects with localized juvenile periodontitis, advanced destructive periodontitis, or adult periodontitis and in normal persons. Significantly elevated serum immunoglobulin G (IgG) antibody levels to B. gingivalis were seen in adult and advanced destructive periodontitis patients. Serum IgM and IgA antibodies were increased in diseased versus normal subjects, whereas negligible levels of serum IgE antibody were detected to this microorganism. Serum IgG antibody levels to B. intermedius were increased in advanced destructive periodontitis patients; however, the frequency of elevated responses were similar among the groups. Extreme antibody levels to the other Bacteroides spp. were occasionally observed in this population. Additionally, all of the elevated levels were found in diseased patients. Distribution analyses of the antibody levels indicated that most patients exhibited a pattern of elevated antibodies to a limited number of the oral Bacteroides spp. The results suggested that elevated systemic antibody levels to oral Bacteroides spp. are more frequently found in periodontal disease patients. These antibody responses presumably reflect a colonization of the patients. The distribution of the responses may indicate the potential pathogenicity of the microorganisms and is consistent with distinctive host-parasite interactions in this disease.     

Haplotype-based banking of human pluripotent stem cells for transplantation: potential and limitations

High expectations surround the area of stem cells therapeutics. However, the cells' source-adult or embryonic-and the cells' origin-patient-derived autologous or healthy donor genetically unrelated-remain subjects of debate. Autologous origins have the advantage of a theoretical absence of immune rejection by the recipient. However, this approach has several limitations with regard to the disease of the recipient and to potential problems with the generation, expansion, and manipulation of autologous induced pluripotent stem cells (iPS cells) preparation. An alternative to using autologous cells is the establishment of a bank of well-characterized adult cells that would be used to generate iPS cells and their derivatives. In the context of transplantation, such cells would come from genetically unrelated donors and the immune system of the recipient would reject the graft without immunosuppressive therapy. To minimize the risk of rejection, human leukocyte antigen (HLA) compatibility is certainly the best option, and the establishment of an HLA-organized bank would mean having a limited number of stem cells that would be sufficient for a large number of recipients. The concept of haplobanking with HLA homozygous cell lines would also limit the number of HLA mismatches, but such an approach will not necessarily be less immunogenic in terms of selection criteria, because of the limited number of HLA-compatible loci and the level of HLA typing resolution.     

Chlamydia trachomatis infection: host immune responses and potential vaccines

Chlamydia trachomatis causes genital tract infections that affect men, women, and children on a global scale. This review focuses on innate and adaptive immune responses in the female reproductive tract (FRT) to genital tract infections with C. trachomatis. It covers C. trachomatis infections and highlights our current knowledge of genital tract infections, serovar distribution, infectious load, and clinical manifestations of these infections in women. The unique features of the immune system of the FRT will be discussed and will include a review of our current knowledge of innate and adaptive immunity to chlamydial infections at this mucosal site. The use of animal models to study the pathogenesis of, and immunity to, Chlamydia infection of the female genital tract will also be discussed and a review of recent immunization and challenge experiments in the murine model of chlamydial FRT infection will be presented.     

A microculture system for generating haemolytic antibody responses from human tonsillar lymphocytes.

Small numbers of Ficoll-Hypaque purified human tonsillar lymphocytes were stimulated with PWM to produce SRBC-specific PFC in a microculture system. The magnitude of the response varied among different tonsils but was typically between 200 and 1000 PFC/10(6) cells cultured. Little or no response was observed in the absence of PWM. SRBC failed to stimulate a SRBC-specific response and the presence of this antigen in PWM-stimulated cultures depressed the response. The time of the maximum response was inversely related to the number of cells cultured. In addition, the duration of the response was limited by rapid depletion of critical medium requirements and/or build up of inhibitory factors especially when the cell concentration exceeded 5 x 10(5) cells/culture. This effect could be partially overcome by daily feeding of cultures with fresh medium. Fractionation studies indicated a requirement for both T and B cell populations. Constant efficiency of PFC production with respect to cell number could be achieved by the addition of inactivated autologous 'filler' cells. The significance of these results and applicability of the microculture system to a detailed analysis of human antibody responses will be discussed.     

Parasite-specific antibody and cellular immune responses in human infected with Necator americanus and Oesophagostomum bifurcum

In this study parasite-specific antibody, cellular reactivity and Thl-type or Th2-type cytokine responses were investigated in humans concurrently infected with Necator americanus and Oesophagostomum bifurcum. The prospects for O. bifurcum-specific serodiagnosis based on IgG4 and IgE were evaluated. IgG4 showed low specificity for O. bifurcum due to antigen cross-reactivity with N. americanus, while IgE specifically distinguished between hookworm and O. bifurcum, and, in doubly infected patients, levels of O. bifurcum-specific as well as N. americanus-specific IgE were significantly elevated compared to those with N. americanus mono-infections. Cellular immunity was not strictly dominated by a Thl- or Th2- type reactivity. In co-infected patients cellular unresponsiveness to parasite antigens was observed, while cellular production of tumour necrosis factor alpha (TNF-alpha) and gamma-interferon (IFN-gamma) was greater in those doubly infected. Th2-type cytokines (interleukin-5 and interleukin-10) were produced in equal amounts by peripheral blood mononuclear cells from individuals with mono- and coinfections. Such mixed Thl-type and Th2-type immune responsiveness associated with persisting gastrointestinal parasitic nematodes may reflect a state of infection at which parasite-induced inflammatory and enteropathogenic responses co-exist, and furthermore, helminth coinfection will not only suppress parasite-specific cellular responsiveness but may also direct cytokine production towards a "permissive Th1-type cytokine profile" that favours parasite persistence.     

ELISpot for measuring human immune responses to vaccines

The enzyme-linked immunosorbent spot (ELISpot) assay is one of the most commonly used methods to measure antigen-specific T cells in both mice and humans. Some of the primary reasons for the popularity of the method are that ELISpot is highly quantitative, can measure a broad range of magnitudes of response and is capable of assessing critical cellular immune-related activities such as IFN-γ secretion and granzyme B release. Furthermore, ELISpot is adaptable not only to the evaluation of a variety of T-cell functions, but also to B cells and innate immune cells. It is no wonder that ELISpot has evolved from a research tool to a clinical assay. Recent Phase I and II studies of cancer vaccines, tested in a variety of malignancies, have suggested that ELISpot may be a useful biomarker assay to predict clinical benefit after therapeutic immune modulation. This article will discuss the most common applications of ELISpot, overview the efforts that have been undertaken to standardize the assay and apply the method in the analysis of human clinical trials, and describe some important steps in the process of developing a clinical-grade ELISpot.     

The developing human preterm neonatal immune system: A case for more research in this area

Neonates, particularly those born prematurely, are among the most vulnerable age group for morbidity and mortality due to infections. Immaturity of the innate immune system and a high need for invasive medical procedures in the context of a preterm birth make these infants highly susceptible to common neonatal pathogens. Preterm infants who survive may also suffer permanent disabilities due to organ damage resulting from either the infection itself or from the inflammatory response generated under an oxidative stress. Infections in preterm infants continue to pose important healthcare challenges. Yet, developmental maturation events in the innate immune system that underlie their excessively high vulnerability to infection remain largely understudied. In this review article, we identify pertinent knowledge gaps that must be filled in order to orient future translational research.     

Human secondary immune responses induced by influenza in a serum-free tissue culture system

Peripheral blood lymphocytes are capable of producing antibody in response to antigenic stimulation by a recall antigen, influenza, during culture for 10 days in medium consisting of 1 part Ham's F-12 and 1 part Iscove's modified Dulbecco's medium supplemented with sodium bicarbonate, bovine crystalline insulin, human transferrin, 2-mercaptoethanol, progesterone, and bovine serum albumin. Anti-influenza antibody levels in the supernatants were determined by enzyme-linked immunosorbent assay. Optimal conditions for production of anti-influenza antibody in this serum-free medium were: influenza concentration, 0.032–0.125 HAU/ml; day of harvest, 10; and cell concentration, 3.0–4.0×10⁵ cells in 200 μl of medium per well. Use of serum-free medium will allow examination of the effects of various additives to tissue culture without concern for unknown factors or potential interaction with serum.