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PubMed1985

Detection of cellular and humoral immunity to hepatitis B surface antigen (HBsAg) in asymptomatic HBsAg carriers.

Cell mediated and humoral immunity to hepatitis B surface antigen (HBsAg) was studied in nine asymptomatic HBsAg carriers, nine patients with natural acquired immunity to HB infection and nine HB-susceptible donors. Peripheral T and B lymphocytes from all asymptomatic HBsAg carriers and all HB-immune donors studied were specifically induced into proliferation and anti-HBs secretion when stimulated with low doses of HBsAg (2-30 ng antigen protein/ml) in vitro. This activation was achieved by mixing purified B and/or T cells with optimal concentrations of autologous monocytic cells. T and B cells from the HB-susceptible donors were non-responsive under identical culture conditions. These data do neither substantiate the existence of a qualitative defect in T cell function, nor the absence of circulatory B cells capable of synthesizing anti-HBs in vitro in asymptomatic HBsAg carriers. Thus, the inability to mount a satisfactory antibody response to HBsAg in vivo might be a consequence of altered immune responsiveness to this antigen, which may be a relevant factor in the pathogenesis of asymptomatic HBsAg carriership.

Adult↗

PubMed1986

Simple sugars with affinity for the macrophage asialoglycoprotein receptor are adjuvants for the humoral immune response to neuraminidase-treated sheep erythrocytes.

The influence of monosaccharides on the humoral immune response of mice to normal and neuraminidase-treated sheep red blood cells (SRBC) was investigated. In these studies, both the sugars and antigen were administered i.p. D-Galactose displayed adjuvant activity for neuraminidase-treated but not for normal SRBC. This activity was optimal at an antigen dose of 3 X 10(6) cells. Other monosaccharides with an axial hydroxyl group at position 4 and D-mannose-6-phosphate behaved like D-galactose, whereas structurally unrelated sugars did not. The adjuvant activity of the saccharides corresponded to the affinity of the substances for the asialo-glycoprotein receptor. The adjuvant effects were also expressed in cobra venom factor-treated and C5-deficient mice. This suggests that blockade of the asialo-receptor rather than complement is involved in this form of immunoadjuvant activity. The findings are also in support of our hypothesis that obstruction of nonimmune antigen elimination, in this case at the level of macrophages, would be one of the mechanisms underlying immunologic adjuvant activity.

Adjuvants, Immunologic↗

PubMed1984

Humoral immune responses in murine pregnancy. IV. Strain dependency and alloantibody specificity.

The relationship between the ability of female inbred mice to produce an anti-paternal humoral immune response to allogeneic multiparity and the genotype of the female and male strains has been investigated. Only three, all H-2b haplotype strains, were 'responder' strains and produced anti-paternal alloantibody which did not exhibit C'-dependent cytotoxicity. 'Non-responder' strains produced no alloantibody in spite of multiple pregnancies with H-2 and non-H-2 incompatible male strains. However, even responder strains did not produce alloantibody with all incompatible male strains. The absence of a response in a responder strain mated with a male strain differing at only the H-2 locus implicated a role for non-H-2 influences. A study of the specificity of pregnancy-induced alloantibody suggested that this represented only a fraction of the total alloantibody population induced by conventional immunization in the same strain combination. It is suggested that in pregnancy only the anti-non-H-2 humoral responses remain similar to those induced by immunization, whereas anti-H-2 humoral responses are either absent or are restricted to a fraction of the total foreign H-2 specificities presented. These observations are discussed in relation to the nature of the immunogenic stimulus in pregnancy and the expression of histocompatibility antigens on the feto-placental unit.

Animals↗

PubMed2000

Control of LMP7 expression in human endothelial cells by cytokines regulating cellular and humoral immunity.

Formation of antigenic peptides by the multicatalytic proteinase complex (MPC, proteasome) is facilitated by incorporation of three subunits (LMP2, LMP7 and LMP10) that are inducible by IFN-gamma and TNF-alpha. These cytokines, or their functional homologues (e.g. TNF-beta), are released from many cells including Th(1)lymphocytes. To learn more about the relationship between control of cellular immunity and expression of LMP subunits, we measured LMP7 levels in human umbilical vein endothelial cells of cytokines promoting cellular immunity (IL-12, IFN-gamma, TNF-alpha) or humoral immunity (IL-10, IL-6). Little or no effect was seen when cells were exposed to IL-6, IL-10 or IL-12 alone. IFN-gamma upregulated LMP7 levels, as did TNF-alpha to a lesser extent. IL-10 downregulated IFN-gamma-induced increases in LMP7 levels, as did IL-12. The findings indicate that regulation of levels of LMP7 is similar to and may be coupled with that of other molecules required for MHC class I-dependent immunity, and depends primarily on cytokines released by Th(1)helper lymphocytes.

Blotting, Western↗

PubMed2000

Humoral immune responses to DNA vaccines expressing secreted, membrane bound and non-secreted forms of the Tania ovis 45W antigen.

The antibody response to DNA vaccines expressing secreted, membrane bound and non-secreted forms of the same antigen was investigated. The antigen gene selected for these studies was the full length 45W antigen gene from Taenia ovis. This gene encodes a host protective membrane bound antigen with a native secretion signal at the amino terminus and a hydrophobic anchor domain at the carboxyl terminus. Full length and rationally truncated forms of the 45W antigen gene were generated and used to construct DNA vaccines encoding membrane bound, secreted and non-secreted forms of the 45W antigen. The cellular localisation of these antigen forms was confirmed by Western blot studies. BALB/c mice were immunised intramuscularly with plasmid DNA and serum antibody responses measured by enzyme linked immunosorbant assay (ELISA). The cellular localisation of DNA vaccine antigen had a significant effect on the magnitude but not the subclass of antibody responses. Immunisation with DNA expressing secreted 45W generated three-fold higher antibody titres than immunisation with DNA expressing membrane bound 45W, and 18-fold higher antibody titres than DNA expressing non-secreted 45W. All mice generated a predominantly IgG1 antibody response indicative of a TH-2 type immune response. These results indicate that the optimal induction of humoral immune responses to intramuscular genetic immunisation with the 45W antigen, requires the active secretion of antigen. This observation may be of value during the design of DNA vaccines in the future.

Animals↗

PubMed1992

Dilated cardiomyopathy with special reference to humoral immunity.

The question of whether the etiology of DCM is immune or autoimmune has been increasingly discussed. Abnormal findings on humoral immunity in DCM were investigated, especially those regarding anti-heart antibodies (AHA), IgG subclasses and soluble interleukin-2 receptor (sIL-2R). The heterophile type AHA was detected in 64.7% of cases by the indirect immunofluorescence technique (IF) with rat heart, by indirect IF with human heart AHA in 57.8% of cases, and by thin-layer chromatogram with human glycolipids AHA in 44% of cases. Also, 57.1% of the specimens were found to bind IgG on perimyocytes by direct IF with biopsy specimens taken from patients with DCM. The epitope of an antigen which reacted with the heterophile type AHA is a Gal alpha 1-3Gal structure. 200 Kd, 70 Kd and 40 Kd antigens were reacted with AHA detected by indirect IF with human heart. The possible mechanisms of AHA in the pathogenesis could be either complement dependent cytotoxicity or interference to cardiac metabolism. The concentration of sIL-2R and IgG3 in sera from patients with DCM were elevated. These results suggest that immunological abnormalities occur continuously in DCM.

Animals↗

PubMed1995

Humoral immune responses to systemic Candida albicans infection in inbred mouse strains.

The protective role of humoral antibodies in the resolution of systemic candidiasis remains controversial. Investigation of the humoral immune responses in mouse strains of varying susceptibility to infection may demonstrate a link between mouse strain susceptibility, antibody production and specificity, and the ability to resolve an infection. The antibody response in five different strains of mice during primary immune response to systemic infection with Candida albicans was investigated. Immune sera were fractionated by protein A affinity chromatography to yield fractions containing IgG1, IgG2a and IgG2b immunoglobulins. BALB/c mice of low susceptibility to the infection and DBA/2J mice of high susceptibility produced increased levels of the IgG1 isotype and decreased levels of the IgG2a isotype. AKR, CBA/H and C57B1/6J mice of moderate susceptibility produced antibodies predominantly of the IgG2a isotype. The patterns of antigen recognition by antibodies in immune sera and in fractions obtained after protein A chromatography of immune sera were investigated by western blotting and immunostaining. Antibodies from AKR(H-2K) and CBA/H (H-2k) mice reacted strongly after immunoblotting with antigens of 87 and 96 kDa. In contrast, immune sera from both the highly susceptible DBA/2J (H-2d) mice and the resistant BALB/c (H-2d) mice reacted strongly with an antigen of 48 kDa. C57B1/6J (H-2b) mice produced variable antibody reactivity to antigens of 48, 65, 66 and 79 kDa depending on the IgG subclass tested. The IgG subclass responses and the patterns of antigen recognition in these mice suggest that humoral responses to C. albicans may be restricted by H-2 haplotype.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1998

Comparative analysis of humoral immune responses to HIV type 1 envelope glycoproteins in mice immunized with a DNA vaccine, recombinant Semliki Forest virus RNA, or recombinant Semliki Forest virus particles.

The Semliki Forest virus (SFV) system seems to be a useful new approach for generating effective immune responses against HIV-1 in animal models. We evaluated this system by comparing the humoral immune responses raised in mice immunized against the HIV-1 envelope with the SFV system, a DNA vaccine, and a recombinant Env glycoprotein. gp160 ELISA antibody titers (204,800) were highest in the sera from mice immunized with recombinant Semliki Forest virus particles. These sera contained antibodies to the CD4-binding site and recognized linear epitopes on gp120 and gp41 that were also recognized by a pool of sera from HIV1-infected individuals. This demonstrates that the HIV-1 envelope produced in vivo by the SFV system does not fold aberrantly. A low level of neutralizing antibodies against the HIV-1LAI strain was also detected in the serum of one mouse immunized with recombinant SFV particles, suggesting that booster injections should be given to achieve a more effective immune response. SFV recombinant particles induced the strongest humoral responses to the HIV-1 envelope of all the potential HIV env vaccines tested.

AIDS Vaccines↗

PubMed1995

Humoral immune responses to Theileria parva in cattle as measured by two-dimensional western blotting.

Humoral immune responses to schizont antigens from six stocks of Theileria parva were compared by two-dimensional Western blotting using sera from cattle that had been infected with a T. parva stock or a clone. Isoelectric points of a polymorphic immunodominant molecule (PIM) of schizonts that induces strong antibody responses in cattle ranged from acidic to basic. Molecular masses (Mr) of the PIM of the respective T. parva stocks were as follows: T. parva Muguga, 86 kDa; Mariakani, 83 kDa; Marikebuni, 83 kDa; Uganda, 83 kDa; T. parva Boleni, 83 kDa; and T. parva 7014, 100 kDa. Among nine cattle infected with T. parva Muguga, four produced antibodies to a basic antigen having an Mr of 32 kDa. The PIM of T. parva Muguga, T. parva Boleni, and T. parva 7014 reacted strongly with serum obtained from an animal that had been infected with T. parva Muguga. Two-dimensional Western blotting using antischizont monoclonal antibodies enabled us to differentiate between stocks of T. parva.

Animals↗

PubMed1996

Humoral immune responses in Gambians to Pfs16, an immunodominant, Plasmodium falciparum integral membrane protein.

Naturally acquired humoral immune responses to Pfs16, an integral membrane protein expressed in Plasmodium falciparum gametocytes and sporozoites, were investigated in The Gambia. A high prevalence of antibodies to this molecule was detected by peptide ELISA. Ninety-three per cent of the people taking part in a survey at the end of the rainy season (November) had serum antibodies to one or more synthetic peptides spanning the sequence: 88% reacted with one particular peptide sequence (IMLIILSGIVGFKVK) whereas only one out of ten non-Gambians (taking anti-malarial prophylaxis with no history of infection) reacted with the peptide. Epitope mapping with mouse MoAbs has shown that this peptide is located on a part of the molecule differing from immunodominant regions of the molecule identified in a previous study in Papua New Guinea.

Adolescent↗

PubMed2005

Diverse humoral immune responses and changes in IgG antibody levels against mycobacterial lipid antigens in active tuberculosis.

Humoral immune responses of active TB patients against six mycobacterial lipid antigens [trehalose 6,6'-dimycolate (TDM) from Mycobacterium bovis BCG (TDM-T) and Mycobacterium avium complex (TDM-M), trehalose 6-monomycolate (TMM) from M. bovis BCG (TMM-T) and M. avium complex (TMM-M), triacyl (PL-2) and tetraacyl (PL-1) phosphatidylinositol dimannosides] were examined by ELISA. IgG antibodies of TB patients with active disease reacted against the six lipid antigens distinctively, but heterogeneously. If tests were combined and an overall positive was scored cumulatively when any one of the six tests was positive, a good discrimination between patient and normal subject was obtained. A positive result in any one of the six tests was obtained in 91.5% of all 924 hospitalized patients and 93.3% of 210 patients at their first visit to the outpatient clinic. The IgG antibody response differed considerably from patient to patient, and the response patterns were grouped into several types. IgG antibody levels paralleled the bacterial burden; however, the smear-negative (culture-positive) patient group also showed high positive rates and mean ELISA DeltaA values against the six lipid antigens. There were also marked differences in positive rate and mean DeltaA values between cavity-positive and -negative groups, the former being higher than the latter. After anti-TB chemotherapy was initiated, IgG antibody levels decreased dramatically, paralleling the decrease in the amount of excretion of bacteria. Since multiple-antigen ELISA using particular lipid antigens was highly sensitive, and IgG antibody levels vary greatly at different stages of the disease, this technique is applicable for early diagnosis of smear-negative (and -positive) active TB and the prognosis for completion of anti-TB chemotherapy.

Adult↗

PubMed1981

Decreased in vitro humoral immune responses in aged humans.

Induction of antigen-specific and non-specific (polyclonal) humoral immune responses in vitro was investigated in peripheral blood mononuclear cells of aged (65-85 yr) and young (20-30 yr) volunteers. In vitro immunization of lymphocytes with antigen (sheep erythrocytes) was performed in a recently described microculture system, and anti-sheep erythrocyte plaque forming cells were quantitated in a direct hemolytic plaque assay. Immunoglobulin secreting cells, induced polyclonally with pokeweed mitogen, were quantitated in a reverse hemolytic plaque assay. Significant depressions of antigen-specific as well as polyclonal responses were noted in relation to advancing age. Antigen-specific responses were more frequently depressed than polyclonal responses. T cell mitogen concanavalin A (Con A) was used to amplify functions of autologous immunoregulatory T cells. Addition of 10 microgram/ml Con A to lymphocytes of young donors at culture initiation resulted in activation of suppressor cells and abrogated antigen-specific responses. Delayed addition of Con A, on the other hand, enhanced responses, presumably because of activation of helper T cells. Similar manipulations of lymphocyte cultures from aged donors showed failure of Con A to suppress antigen-specific responses in approximately half of the responders. In many nonresponders, responses within normal range were elicited by the delayed addition of Con A to their lymphocyte cultures. Deviations beyond the range of expected responses were noted in 32.5% of the co-cultures between pokeweed mitogen stimulated young and aged cells. Our findings suggest that age-related deficiencies of B cell function are frequently associated with dysfunction of immunoregulatory T cells and are only occasionally due to intrinsic defects of B cells.

Adult↗

PubMed1995

Characterization of the humoral immune response to porcine reproductive and respiratory syndrome (PRRS) virus infection.

The development of the humoral immune response against porcine reproductive and respiratory syndrome (PRRS) virus was monitored by an indirect fluorescent antibody (IFA) test, immunoperoxidase monolayer assay (IPMA), enzyme-linked immunosorbent assay (ELISA), and serum virus neutralization (SVN) test over a 105-day period in 8 pigs experimentally infected with ATCC strain VR-2402. Specific antibodies against PRRS virus were first detected by the IFA test, IPMA, ELISA, and the SVN test 9-11, 5-9, 9-13, and 9-28 days postinoculation (PI), respectively, and reached their maximum values by 4-5, 5-6, 4-6, and 10-11 weeks PI, respectively, thereafter. After reaching maximum value, all assays showed a decline in antibody levels. Assuming a constant rate of antibody decay, it was estimated by regression analysis that the ELISA, IFA, IPMA, and SVN antibody titers would approach the lower limits of detection by approximately days 137, 158, 324, and 356 PI, respectively. In this study, the immunoperoxidase monolayer assay appeared to offer slightly better performance relative to the IFA test, ELISA, and SVN test in terms of earlier detection and slower rate of decline in antibody titers. Western immunoblot analysis revealed that antibody specific for the 15-kD viral protein was present in all pigs by 7 days PI and persisted throughout the 105-day observation period. Initial detection of antibodies to the 19-, 23-, and 26-kD proteins varied among pigs, ranging from 9 to 35 days PI. Thereafter, the antibody responses to these 3 viral proteins of PRRS virus continued to be detected throughout the 105-day study period.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed2002

Humoral immune response to an allogenic foetus in normal fertile women and recurrent aborters.

BACKGROUND: Several humoral immune factors are responsible for a successful pregnancy. There are a few studies, which demonstrate the role of antipaternal cytotoxic antibodies (APCA) and mixed lymphocyte reaction blocking factor (MLR-Bf) in the maintenance of pregnancy. However, these factors have not been studied in women with history of recurrent spontaneous abortion (RSA). We designed this study to review the role of APCA and MLR-Bf in normal pregnancy as well as in women with a history of RSA. METHODS: One hundred and five women with unexplained recurrent spontaneous abortion were included in the present study. These women were screened for all other known causes of recurrent abortion. We also included 60 normal fertile women, fifteen from each trimester and fifteen women during the post partum period (up to six months). RSA and controls (normal pregnancy) were matched for age, caste, and socio economic background and also for parity. APCA and MLR-Bf were evaluated in all the groups. All women with RSA who conceived during the study period were on follow up. RESULTS: We have analyzed the status of APCA and MLR-Bf in normal pregnancy (different gestational periods and during post partum), and in women with history of RSA. Our results show that APCA was significantly higher in controls as compared to RSA women. MLR-Bf was directed against the husbands' cells in normal pregnancy and was virtually absent in RSA women. CONCLUSION: Our results indicate that there is a significantly low titer of APCA and MLR-Bf in women with recurrent spontaneous abortion. This highlights the role of these factors in the maintenance of successful pregnancy.

Journal Article↗

PubMed1976

Analysis of the humoral immune response to influenza virus in vitro.

The induction of in vitro primary and secondary humoral immune responses to influenza virus in murine splenic explant culture is described. Anti-influenza antibody synthesized in vitro was detected and quantitated by a radioimmunoassay which utilized influenza coupled to bromoacetylcellulose. Both in vitro primary and secondary responses could be stimulated over a large range of virus doses. In vitro secondary responses were maximal when spleen donors had been immunized by the parenteral route although secondary type responses could be demonstrated as well after primary immunization by pulmonary infection. In vitro stimulation with influenza virus was relatively insensitive to inhibition at high antigen doses. The results are discussed in terms of the response of other antigens in spleen fragment culture.

Animals↗

PubMed1991

A biorhythm in the humoral immune response to SRBC in mice.

We reveal an existence of a biorhythm in the humoral immune response to SRBC in vivo and in vitro, basing on our experimental data registered in the years 1978-1991. It appears that the immune response to SRBC undergoes regular cycles of high and low responsiveness with 4-week intervals. The biorhythm is not dependent on sex or strain of mice. The importance of the phenomenon in the laboratory practice is discussed.

Animals↗

PubMed1990

[The development of the epidemic process and the formation of humoral immunity to the hepatitis A virus in adolescent collectives].

Additional observation on the formation of humoral immunity to hepatitis A virus has been carried out in 2,375 adolescents aged 15-17 years and belonging to 12 groups at 3 areas of the USSR. This observation has shown that in the presence of a high level of the immune stratum the spread of infection occurs, as a rule, outside the group under observation. Besides, as revealed in this study, the risk of seroconversion and the loss of specific antibodies are, respectively, directly and inversely related to the level of the immune stratum in the group. The level of the immune stratum in a given age group supposedly reflects the intensity of the development of the epidemic process at a definite area.

Adolescent↗

PubMed1978

Humoral immune responses of cattle vaccinated with leptospiral pentavalent outer envelope and whole culture vaccines.

The humoral immune response of cattle vaccinated with leptospiral pentavalent outer envelope and whole culture vaccines was monitored with the microscopic agglutination (MA) test and the leptospiricidal activity (LA) test for a 2.5-year period. The serovars present in the vaccines were canicola, icterohaemorrhagiae, grippotyphosa, pomona, and hardjo. The LA test had greater sensitivity than the MA test for detecting antibodies to the hardjo, canicola, and icterohaemorrhagiae components of the pentavalent vaccine and was approximately equal in sensitivity to the MA test for detecting antibodies to the grippotyphosa and pomona components of the leptospiral vaccine. Both IgM and IgG were produced in similar amounts in the immunized cattle, and both classes were reactive in the MA and the LA tests. The IgM class of antibodies was more reactive in the MA test, whereas the IgG class of antibodies was more reactive in the LA test. Using the MA test as the criterion of evaluating, the immunogenic potency of the individual vaccine components was found to be canicola more than icterohaemorrhagiae more than grippotyphosa more than pomona more than hardjo. A direct correlation between protective antibodies, as determined by the hamster passive protection test, and antibodies reactive in the MA and the LA tests was observed.

Agglutination Tests↗