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Maturation of the humoral immune response in mice.

In spite of the prenatal appearance of immunoglobulin-bearing lymphocytes and theta-positive lymphocytes in the spleens of Swiss-L mice, these mice are not able to produce detectable levels of humoral antibodies in response to antigen until after 1 wk of age. Adult levels of response are not achieved until 4-8 wk of age. In the presence of bacterial lipopolysaccharides, which can substitute for or enhance T-cell function, the B cells from young Swiss-L mice were found to be indistinguishable in function from adult B cells, both with respect to the numbers of plaque-forming cells (PFC) produced in vitro in response to antigen and with respect to the kinetics of PFC induction. The spleen cells from young Swiss-L mice are significantly less sensitive than adult spleen cells, however, to stimulation by the T cell mitogens, concanavalin A (Con A) and phytohemagglutinin (PHA). Very few Con A-responsive cells could be detected at birth but the numbers increased sharply with age until 3 wk after birth. On the other hand, PHA-responsive cells could not be detected in the spleen until about 3 wk of age. The latter cells were found to respond also to Con A, but at a lower dose (1 microg/ml) than that required for the bulk of the Con A-responsive cells (3 microg/ml). The cells that respond both to PHA and to Con A appear in the spleen at about the time that Swiss-L mice acquire the ability to produce humoral antibodies, and these cells can be depleted from the spleen by the in vivo administration of antithymocyte serum. The development of humoral immune responses in these mice therefore appears to be correlated with the appearance of recirculating T lymphocytes that are responsive both to PHA and to Con A.

Age Factors↗

[Estradiol-17beta, sex steroid-binding globulin and the cellular and humoral immunity indices in young women with breast cancer].

Blood serum levels of sex steroid-binding globulin (SSBG), free and SSBG- and albumin-binding fractions of estradiol-17 beta as well as some indexes of cellular and humoral immunity were compared in 25 young female patients with stage 1-3 breast cancer and controls. No significant difference in SSBG and free estradiol-17 beta fraction as well as free estrogen index was established between patients and controls. In the patient group, a direct correlation was established between free estradiol-17 beta fraction level and that of mature T-cells expressing CD3 antigen and between SSBG level, on the one hand, and total level of T-cells (CD7 antigen) and that of T-suppressors (CD8 antigen-containing cytotoxic lymphocytes), on the other. An inverse correlation was established in the same group between SSBG level and that of B-cell markers expressing CD3 and 7 antigens and IgM mu-chain. In the follicular and lutein phases of the menstrual cycle, a difference in the degree of correlation between total estradiol-17 beta level and immunologic indexes was established between breast cancer patients and controls.

Adult↗

Diminished expression of CD40 ligand may contribute to the defective humoral immunity in patients with MHC class II deficiency.

Major histocompatibility complex (MHC) class II deficiency (bare lymphocyte syndrome, BLS) is a rare primary immunodeficiency classified as a subgroup of severe combined immunodeficiency. We studied T and B lymphocyte function by examining the CD40 ligand/CD40 system in three BLS patients from two unrelated families. CD40 ligand expression by maximally activated BLS T cells was diminished. This abnormality may represent immunological naïveté rather than a general T cell defect, since expression of activation marker CD69 and proliferative responses to PHA or anti-CD3 were normal, and BLS T cells primed and restimulated in vitro expressed normal amounts of CD40 ligand. BLS B cells proliferated and produced IgE if stimulated with anti-CD40 or soluble CD40 ligand and IL-4. Activation of BLS B cells with soluble CD40 ligand and IL-4 induced normal expression of activation markers, although MHC class II expression remained absent. Depressed antibody titers, lack of amplification and failure to undergo isotype switching in response to immunization with bacteriophage phi x 174 demonstrated defective T cell help. We conclude that BLS B cells are functionally normal if appropriately stimulated, and that the defective humoral immunity observed may be related to diminished expression of CD40 ligand on BLS T cells.

Antibodies, Viral↗

Humoral immune responses to T cell tropic retrovirus simian T lymphotropic virus type III in monkeys with experimentally induced acquired immune deficiency-like syndrome.

The T cell tropic retrovirus of macaque monkeys simian T lymphotropic virus type III (STLV-III) has morphologic, growth, and antigenic properties indicating that it is related to human T cell lymphotropic virus type III/lymphadenopathy-associated virus (HTLV-III/LAV), the etiologic agent of the acquired immune deficiency syndrome (AIDS) of humans. STLV-III has recently been shown to induce an AIDS-like disease in macaque monkeys. In this study the humoral immune responses of six experimentally infected monkeys have been characterized to determine whether certain parameters of the antibody response to the virus might be predictive of the clinical outcome of this infection. Two distinct patterns of antibody responses were found. Four animals that died within 160 d of inoculation developed low titer anti-STLV-III antibody responses that recognized only the viral envelope protein, and progressive declines in total plasma IgG levels and absolute peripheral blood T4 lymphocyte numbers. The two animals that lived longer (one died at 352 d, the other remains alive at 430 d) developed high titer anti-STLV-III antibody responses that recognized both viral envelope and core proteins, increases in total plasma IgG, and a later decrease in number of peripheral blood T4 lymphocytes. Interestingly, the single animal that has remained clinically healthy after infection was the only one to develop detectable STLV-III neutralizing antibodies.

Acquired Immunodeficiency Syndrome↗

Cellular and humoral immune responses to varicella-zoster virus in immunocompromised patients during and after varicella-zoster infections.

Humoral and cell-mediated immune responses to varicella-zoster (V-Z) virus were assessed in patients during and after V-Z infections. Ongoing V-Z infections was associated with minimal cellular immunity but not necessarily with poor humoral immunity. Recovery from V-Z infection was associated with a vigorous cellular immune response. Cell-mediated immunity to V-Z virus was demonstrable for years after varicella, but responses were lower in immunocompromised patients than in normal individuals.

Adolescent↗

Preferential humoral immune response in prostate cancer to cellular proteins p90 and p62 in a panel of tumor-associated antigens.

BACKGROUND: Cytoplasmic p90 autoantigen was recently cloned from a cDNA expression library using serum antibody from a cancer patient. The humoral immune response to p90 in prostate cancer and benign prostatic hyperplasia (BPH) was examined. METHODS: An antigenic fragment of recombinant p90 protein and several other tumor-associated antigens (TAAs) were used in ELISA and Western blotting to detect antibodies in sera from patients with prostate cancer, BPH, and other controls. RESULTS: Autoantibodies to p90 were detected in 30.8% of 133 prostate cancer patients versus 1.5% in 68 BPH patients. When a selected panel of six TAAs including p90 were used for immunoscreening, the cumulative positive reactions in prostate cancer sera reached 92.5%, significantly higher than in BPH and other control sera. Antibodies to p90 showed the highest frequency in prostate cancer (30.8%), followed by antibodies to p62 (22.6%). CONCLUSIONS: A panel of six selected TAAs was shown to have high sensitivity and specificity as immunodiagnostic markers in prostate cancer.

Antigens, Neoplasm↗

Head and neck cancer antigens recognized by the humoral immune system.

Head and neck cancer in advanced stages are difficult to treat. Therefore, development of new treatment modalities and preventive measures are required. We now report the identification of human head and neck cancer antigens recognized by the humoral immune system. We used the serological analysis of recombinant cDNA expression libraries (SEREX) approach. cDNA libraries from cell lines of squamous cell carcinoma of head and neck (SCCHN) and a normal testicle tissue were screened using sera from six allogeneic SCCHN patients. Total 28 positive clones belonging to 19 different genes were identified, including 12 known genes and 7 unknown ones. Expression analysis on 13 normal tissues and 13 cancer tissues using reverse transcription-PCR (RT-PCR) revealed eight ubiquitously expressed genes, nine of which were expressed preferentially in cancer tissues and two cancer/testis antigens. These antigens we defined may be pertinent candidate antigens for future cancer-diagnosis and related immunotherapy.

Antibody Formation↗

Immunoblot analysis of the humoral immune response in primary cytomegalovirus infection.

Cytomegalovirus is a common infection in immunologically normal adults. It may cause an asymptomatic infection or may manifest symptomatically as heterophilic-negative mononucleosis. Studies of CMV infection in immunocompromised patients indicate that humoral immune response plays a role in modulating disease severity; however, the effect of antibodies to CMV in modifying disease expression and transmission in immunologically normal individuals has not been well characterized. Using immunoblot technology, we have demonstrated that immune serum from normal adults contains antibodies to at least 15 CMV-associated proteins and that there is strain-to-strain variation in the expression of these immunogens. The kinetics of the immune response were evaluated by using serial sera collected from normal adults after primary CMV infection; analysis of immunoblots of these sera identified one group of antibodies to CMV proteins that arise early and are stable over time, a second group that appear late after infection, and a third group that are variable among patients and over time.

Adult↗

Visceral schistosomiasis of domestic animals in India: humoral immune status of infected cattle, sheep and goats against major polypeptide antigens of Schistosoma indicum and S. spindale.

Polypeptide profiles of Schistosoma indicum and S. spindale adult worm homogenates were obtained by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Humoral immune status of infected cattle, sheep and goats against Schistosoma indicum and S. spindale Ags was determined by immunoblot analysis and by indirect ELISA using four major polypeptides of approximate molecular masses 45 kDa, 40 kDa, 28 kDa and 15 kDa electro-eluted from the gel slices. Cattle sera samples had higher levels of antibodies against Si/s40 and Si/s28 than against Si/s45 antigen. Reasons have been discussed for the absence of detectable levels of anti-Si/s28, -Si/s45 and -Si/s40 antibodies in a significant number of sera samples from S. indicum egg-positive sheep.

Animals↗

[Genetic control of humoral immune response to xenografts. A monoclonal antibody that causes the hyperacute rejection of cardiac xenografts uses the genes in a native form].

The early phases of the rejection of xenografts exchanged between closely-related species are dominated by a vigorous humoral immune response. We have recently used a linkermediated polymerase chain reaction to generate Ig heavy and light chain specific cDNA libraries to examine the Ig gene control of a prototypic IgM monoclonal antibody, HAR-1, that causes the hyperacute rejection of hamster xenografts. Recombinant clones from the library were screened directly from bacterial colonies by PCR and the nucleic acid sequences of the clones established. Our results demonstrate that the HAR-1 hybridoma is encoded by Ig VH and JH genes in a germline configuration. Comparison of the cDNA sequence for HAR1-VH with the germline equivalent of the gene isolated from newborn LEW liver (provisionally designated VH1.1) showed that the two VH sequences share a nucleic acid identity of 99,3%. Similarly, the HAR-1 monoclonal uses a Ig JH gene that is 98,2% identical with the JH1 nucleic acid sequence available in the GenBank. The use of Ig VH and JH genes in a germline configuration is similar to that seen with polyreactive natural antibodies to infectious agents and autoantibodies. These humoral responses are thought to be the results of the stimulation of a T cell-independent subset of B cells, the B-1a/B-1b subset, that are responsible for producing antibodies that serve as a primitive humoral (natural antibody) defense mechanism against infectious diseases. Our results suggest that the humoral component of the rejection of xenografts in the hamster-to-rat model may represent the stimulation of this type of B cell antibody response by xenogeneic target antigens that share antigenic epitopes with bacteria and other infectious agents.

Animals↗

Our current understanding of humoral immunity of poultry.

With the inception of the modem era of avian immunology in the mid-1950s following the discovery of the role of the bursa of Fabricius (BF), steady and persistent research has been conducted by scientists. Findings that have advanced our knowledge of humoral immunity (HI) include the distinct role of B cells in antibody production, explanation of antibody diversity by gene rearrangement and conversion, identification of a bursal secretory dendritic cell and dendritic cells of the spleen, characterization of cytokine factors, and mapping of MHC class II genes. Numerous experiments have been conducted to evaluate the HI responses of poultry through antibody titer determination and quantitation of immunoglobulin concentrations in the humors. Antibody studies have included the genetics of HI regarding both quantity and persistence of responses as influenced by multiple genes and MHC haplotypes. In the past decade, emergence of monoclonal antibody reagents to identify cell surface markers on B cells and accessory cells has allowed for discrimination of HI functions of cells by immunohistochemistry, flow cytometry, and other techniques. The signaling mechanisms of avian B cells are becoming better understood. Continued work in avian immunity has led to more advances in identification, characterization, and gene sequencing of important cytokines. Additional work is required to identify and sequence genes for many more cytokines that have direct effects on B cells. A brief history of research published on avian B cells is presented with an examination of some of the important research findings reported in recent years.

Allergy and Immunology↗

Transient siRNA-mediated attenuation of liver expression from an alpha-galactosidase A plasmid reduces subsequent humoral immune responses to the transgene product in mice.

Hepatocytes are an effective depot for protein production from gene therapy vectors. However, when gene transfer vectors or their delivery induces hepatic inflammation, adaptive immune responses against the transgene product can ensue. In BALB/c mice, hydrodynamic delivery of a CMV-driven plasmid DNA (pDNA) bearing human alpha-galactosidase A (alphagal) to the liver generated antibodies against alphagal. This humoral immune response was more robust in a transgenic knockout for alphagal, the Fabry mouse. The antibody response could be attenuated in both mouse strains by using a promoter more restricted to hepatocytes. In an attempt to reduce further the humoral responses to alphagal, expression from the transgene was attenuated by using siRNA during the period of initial delivery-associated liver inflammation. In both mouse models and with both promoters, codelivering an alphagal siRNA resulted in a 2 log decrease in initial expression that then increased over the next few weeks to levels generated by the pDNA alone. This strategy led to both attenuated antibodies and an immune status approximating "tolerance" to alphagal. Importantly, in the Fabry mouse, an alphagal siRNA together with a hepatocyte-restricted promoter gave minimal anti-alphagal antibodies and profound tolerance, suggesting that such an approach might have clinical utility for genetic diseases.

Animals↗

Suppression of cellular and humoral immunity to T-dependent antigens by calorie restriction.

Calorie restriction is known to preserve the immunologic function and prolong the life span of autoimmune-susceptible mice. In order to determine the influence of calorie restriction on cellular and humoral immunity, we tested lymphocyte proliferative response and antibody response to T-dependent antigens in C57Bl/6 mice maintained on restricted diets. C57Bl/6 mice fed a low-calorie diet demonstrated a marked reduction in T-dependent-antigen-specific lymphocyte proliferation and antibody response when compared to mice fed ad libitum. The depressed lymphocyte response seen in calorie-restricted animals is attributed to a defect in both the macrophages and T cells in antigen processing, presentation, and/or proliferation.

Animals↗

Organophosphorus pesticide immunotoxicity: effects of O,O,S-trimethyl phosphorothioate on cellular and humoral immune response systems.

The time course of immunosuppression induced by acute treatment with O,O,S-trimethyl phosphorothioate (OOS-TMP), an impurity in technical formulations of malathion, was examined in female C57B1/6 mice. Both cell-mediated and humoral immune responses were examined and included allospecific cytotoxic T cells, proliferative response to mitogens, interleukin-2 production and antibody production to sheep red blood cells. OOS-TMP pretreatment led to a reversible suppression of the generation of cytotoxic T lymphocytes and antibody-secreting cells to sheep erythrocytes. However, the mitogenic response of splenocytes from animals treated with nontoxic doses of OOS-TMP (as measured by body weight loss, serum cholinesterase levels and splenic lymphocyte number) to concanavalin A was not significantly suppressed, but the response to the B cell mitogen lipopolysaccharide was slightly decreased on day 1 following treatment. In contrast, interleukin-2 production was elevated by 24 h following treatment, but had returned to control levels by day 7. These data suggest that OOS-TMP was able to block the generation of cytotoxic T lymphocytes and antibody responses at doses of OOS-TMP that did not affect body weight or splenic lymphocyte number and this suppression was reversible.

Animals↗

Course of infection and humoral immune reaction in calves infected orally with different Salmonella serovars.

5 groups of five calves each were orally infected with the salmonella serovar S. agona, S. dublin, S. enteritidis, S. infantis or S. typhimurium. The course of infection and the humoral immune response against 5 antigen preparations of the homologous and the 4 heterologous salmonella serovars were monitored in each group. Antibodies against two different protein antigens and against two differently prepared LPS antigens were determined by ELISA and the level of H-agglutinin was assessed by micro agglutination test. All 5 serovars proved to be virulent for calves. The specificity of antibodies produced following the infection are not always consistent with the Kauffmann-White scheme. There was a relation of age and strain for infection on immune response, whereas the severity of infection did no correlate with an increase of antibody production. The serological tests used were good for investigation of a defined host-parasite-interaction but not satisfactory for serovar specific diagnosis of salmonella infection in cattle.

Animals↗

Role of complement receptor type 2 and endogenous complement in the humoral immune response to conjugates of complement C3d and pneumococcal serotype 14 capsular polysaccharide.

Conjugation of the complement fragment C3d to both T-cell-dependent (TD) protein and T-cell-independent type 2 (TI-2) polysaccharide antigens enhances the humoral immune response in mice immunized with either type of antigen. However, the ability of C3d-protein conjugates to enhance the antibody response in mice deficient in complement receptor types 1 and 2 (CR1 and CR2) has raised questions about the role of C3d-CR2 interactions in the adjuvant effect of C3d. In this study, we examined the role of CR2 binding and endogenous complement activation in the antibody response to conjugates of C3d and serotype 14 pneumococcal capsular polysaccharide (PPS14). To block binding of PPS14-C3d conjugates to CR2, mice were immunized with a mixture of vaccine and (CR2)2-immunoglobulin G1 (IgG1). Mice receiving (CR2)2-IgG1 at the time of primary immunization had a marked reduction in the primary anti-PPS14 antibody response but an enhanced secondary anti-PPS14 response, suggesting that C3d-CR2 interactions are required for the primary response but can have negative effects on the memory response. Further, compared with mice receiving PPS14-C3d having a high C3d/PPS14 ratio, mice immunized with PPS14-C3d with low C3d/PPS14 ratios had an enhanced secondary antibody response. Treatment of mice with cobra venom factor to deplete complement had insignificant effects on the antibody response to PPS14-C3d. Experiments with CBA/N xid mice confirmed that PPS14-C3d conjugates retain the characteristics of TI-2 rather than TD antigens. Thus, the adjuvant effect of C3d conjugated to PPS14 requires C3d-CR2 interactions, does not require activation of endogenous complement, and is not mediated by TD carrier effects.

Adjuvants, Immunologic↗

Humoral immunity is not critical for protection against experimental infection with Sarcocystis neurona in B-cell-deficient mice.

Immunodeficient B-cell-deficient mice (mmuMT) were infected with Sarcocystis neurona merozoites to determine the role of B cells and the humoral immune response in protective immunity. As expected, the mice did not seroconvert based on a direct agglutination test. Infected mice did not have significant changes in gross pathology at the time points examined. Histologic changes included mild perivascular and peribronchial infiltrate in the lungs; perivascular infiltrate and mild inflammatory sinusoidal foci in the liver; prominent high endothelial venules in the lymph nodes; and moderate cellular expansion of the periarteriolar sheaths (PALS) in the spleen. Changes resolved by day 60 postinfection. Mice developed significant CD4 and CD8 responses in lymphoid organs, including significant effector (CD45RB(high)) and memory (CD44(high)) CD4 and CD8 responses. Flow cytometry confirmed the lack of B cells. Overall, these data suggest that B cells are not critical to the protective immune response to SN infection.

Agglutination Tests↗

Studies on Dipetalonema viteae (Filarioidea) I. Microfilaraemia in hamsters in relation to worm burden and humoral immune response.

The course of a primary infection with Dipetalonema viteae was studied in one randomly bred and in one inbred strain of hamster. Worm recovery and the duration and intensity of the microfilaraemia were analyzed and related to the humoral immune response of the host by using the indirect immunofluorescent antibody test on frozen sections of female worms, on eggs and on intact microfilariae. The inbred strain showed a greater susceptibility to the parasite. This was evidenced by high worm recovery and prolonged microfilaraemia. The duration of microfilaraemia did not depend on the number of recovered female worms. Most of the randomly bred hamsters suppressed microfilaraemia by week 30 post infection whereas some hamsters of the inbred strain were still microfilaraemic. Splenectomy prior to infection did not affect the duration of microfilaraemia. Antibodies to the cuticle of microfilariae always appeared in the sera after immunity to circulating microfilariae had been built up.

Animals↗