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Humoral immune response against human cytomegalovirus (HCMV)-specific proteins after HCMV infection in lung transplantation as detected with recombinant and naturally occurring proteins.

The humoral immune response to four intracellularly located cytomegalovirus (CMV) proteins was studied in 15 lung transplant recipients experiencing active CMV infections. Five patients had primary infections, and 10 had secondary infections. Antibodies of the immunoglobulin M (IgM) and IgG classes were measured in an enzyme-linked immunosorbent assay (ELISA) system in which procaryotically expressed recombinant proteins were used as a substrate and also in a monoclonal antibody-based capture ELISA which uses naturally occurring proteins as a substrate. The proteins investigated were the lower matrix protein pp65 (ppUL83), the major DNA-binding protein p52 (ppUL44), and the two immediate early proteins IE1 and IE2 (different splicing products of UL123). Higher levels of antibodies were found to pp65 and especially to p52 than to the immediate early antigens. Antibody levels detected in the recombinant protein-based ELISAs were generally lower than antibody responses detected with the matching antigen capture ELISA. Moreover, some patients appeared to have antibodies mainly to epitopes present on naturally occurring proteins. The antibody responses detected in both assays were related to the viral load during infection as assessed by the CMV antigenemia test, which is a quantitative marker for CMV load. It was found that although epitopes on naturally occurring proteins induce higher antibody responses and responses in more patients, antibodies directed to epitopes present on the recombinant proteins were inversely related to the viral load during a CMV infection. Therefore, antibodies to epitopes on the recombinant proteins might be more clinically relevant in this group of lung transplant recipients.

Antibodies, Viral↗

Humoral immune response to chlamydial genital infection of mice with the agent of mouse pneumonitis.

The objective of this study was to characterize the humoral immune response to chlamydial genital infection of mice with the mouse pneumonitis agent (MoPn). With an enzyme-linked immunoabsorbent assay, immunoglobulin G antibodies to MoPn were first detected in plasma by day 14. Peak plasma antibody concentrations were reached by day 49, and this response did not decline significantly throughout the 300-day monitoring period. Immunoglobulin A against MoPn could first be detected in pooled vaginal washes by day 21 after infection and had reached peak concentrations by day 28, but anti-MoPn immunoglobulin G was not consistently present in secretions. The antibody response in secretions had declined slightly by day 300. Immunoblot analysis revealed that the early phase of the plasma antibody response to MoPn as a result of genital infection was against lipopolysaccharide, the major outer membrane protein, and a 62-kilodalton (kDa) protein. In secretions, early-phase immunoglobulin A antibodies were directed to the major outer membrane protein and lipopolysaccharide. Late reactions to 15-, 22-, and 83-kDa proteins in plasma were noted. Late reactions to the 62-kDa protein in secretions were also noted. The cause of these late responses remains unexplained. When mice were challenged intravaginally with MoPn at 50-day intervals after the primary infection, it was found that mice inoculated on day 100 or after were susceptible to reinfection. Susceptibility could not be related to a decline in the antibody concentration in plasma or secretions or in the antibody response to specific components of MoPn as measured by immunoblot analysis.

Animals↗

Longitudinal study of naturally acquired humoral immune responses against the merozoite surface protein 1 of Plasmodium vivax in patients from Rondonia, Brazil.

A longitudinal study on the naturally acquired humoral immune responses against the merozoite surface protein 1 of Plasmodium vivax (PvMSP-1) was performed in malaria patients from the Brazilian Amazon region of Rondonia. We have previously cloned and expressed a recombinant protein, ICB2-5, that encodes 508 amino acids from the N-terminal portion of the PvMSP-1 protein. This affinity-purified polypeptide was tested by an enzyme-linked immunosorbent assay in a one-year longitudinal study using sera from 34 patients who had at least one malaria infection during the study period. The results demonstrated that more than 90% of the sera from patients having experienced more than three previous malaria infections contained antibodies to ICB2-5 at the time of a new clinical episode. Unexpectedly, more than half of these multiple-infected patients had an antibody response to ICB2-5 in which the predominant isotype was IgM. In contrast, more than 83% of the sera from these same patients contained predominantly IgG antibodies against total blood-stage antigen preparations. To determine if these results were due to the lack of boosting against this portion of the PvMSP-1 molecule, the presence of IgG antibodies to ICB2-5 in the sera from 11 patients who had consecutive malarial episodes during the study year was investigated. Five of these eleven patients failed to produce IgG antibodies to ICB2-5 even after 1-3 infections. Thus, these results suggest that no boosting against this region of the PvMSP-1 molecule was achieved by natural infections among these patients.

Adolescent↗

[Diagnostic and prognostic value of the assessment of selected indicators of humoral immunity in alcoholism-related pathology of the liver].

The purpose of the study was evaluation of the usefulness of selected indices of humoral immune responsiveness in the differential diagnosis of post-alcoholic hepatocellular damage. The study was carried out in 105 patients: 10 patients with a history of alcohol abuse without clinical and biochemical evidence of hepatocellular damage, 2) patients with alcoholic cirrhosis, 3) patients with post-inflammatory cirrhosis. The prognostic usefulness of the determinations of serum IgM, C3 and C4 complement components and circulating immune complexes in early diagnostic of alcoholic liver disease was demonstrated. It was noted also that increased serum IgA level may be a useful index differentiating of cirrhosis after hepatitis from alcoholic cirrhosis.

Alcoholism↗

Effects of different levels of dietary selenium and vitamin E on the humoral immunity of rats.

The aim of the present study was to investigate whether the level of selenium and selenium/vitamin E supply influences the humoral immunity of rats. In order to detect the effect of Se supply and age, 36 weaned Sprague-Dawley rats divided into two equal groups were killed after 22 or 45 experimental days by decapitation (Exp. I). In Exp. II 9 groups of 10 rats each were exposed to each combination of deficient, normal or excessive selenium with a vitamin E supply and killed after 44 days. The basic (deficiency) diet which was the same in both experiments contained 0.04mg Se and 8mg vitamin E per kg dry matter. The supplementation per kg diet was 0 or 0.2mg Se and 30mg vitamin E in Exp. I and 0, 0.2 or 1mg Se and 0, 30 or 200mg vitamin E in Exp. II. The concentration of selenium in serum, liver and spleen samples and the activity of glutathione peroxidase, which were determined to define the selenium status of the animals, corresponded well to the required supply situation. The immunoglobulins of type IgA, IgM and IgG with the subtypes IgG1, IgG2a, IgG2b and IgG2c were measured by immunoelectrophoresis. In both experiments selenium deficiency decreased the values of the IgG groups only nominally, IgA was not changed. IgM was significantly reduced, especially with prolonged selenium deficiency and simultaneous vitamin E deficiency. An excessive selenium supply compensated to a great extent for the effects of vitamin E deficiency on IgG and IgA.

Animals↗

Transgenerational priming of immunity: maternal exposure to a bacterial antigen enhances offspring humoral immunity.

Young vertebrates have limited capacity to synthesize antibodies and are dependent on the protection of maternally transmitted antibodies for humoral disease resistance early in life. However, mothers may enhance fitness by priming their offspring's immune systems to elevate disease resistance. Transgenerational induced defences have been documented in plants and invertebrates, but maternal priming of offspring immunity in vertebrates has been essentially neglected. To test the ability of mothers to stimulate the immune systems of offspring, we manipulated maternal and offspring antigen exposure in a wild population of birds, pied flycatchers (Ficedula hypoleuca). We show that immunization of the mother before egg laying apparently stimulates a transgenerational defence against pathogens by elevating endogenous offspring antibody production. If the disease environments encountered by mothers and offspring are similar, this transgenerational immune priming may allow young to better cope with the local pathogen fauna.

Animals↗

Dissection of the humoral immune response toward an immunodominant epitope of HIV: a model for the analysis of antibody diversity in HIV+ individuals.

Understanding the dynamics of the humoral immune response to HIV epitopes in the presence of genetic drift and antigenic variation of the virus may reveal critical elements of protective immunity against HIV. Analysis of antibody maturation and diversity is difficult to study at the molecular level in humans. We used a combinatorial phage display peptide library to elucidate antibody diversity in HIV-infected individuals to a single immunodominant epitope in gp41. A serum sample derived from an HIV+ individual was used to screen a phage display a 12 mer cysteine-constrained loop peptide library. In doing so, we isolated mimotope-presenting phages corresponding to the immunodominant gp41 epitope CSGKLIC (residues 603-609). The mimotopes and control phages expressing epitope variants were reacted with a panel of 30 HIV+ sera. The patients showed distinct and variable recognition patterns compared with one another. Subfractions of the polyclonal sera were affinity purified and analyzed for epitope specificities. These analyses illustrated that epitope variants can be used to decipher antibody diversity. Elucidation of the plasticity of the humoral response and its polyclonality toward discrete epitopes contributes to our understanding of the antibody maturation process in individuals infected with viruses such as HIV.

Amino Acid Sequence↗

Cellular and humoral immune responses of jirds resistant to Dipetalonema viteae infection.

Jirds with prepatent Dipetalonema viteae infections develop an acquired immunity to challenge infections. The objective of the present study was to observe parasite-specific and nonspecific cellular and humoral immune responses in immune jirds. Splenic hyperplasia was observed in infected jirds during the first 5 weeks of infection. Antigen-reactive spleen cells were observed in the lymphocyte transformation assay at 3 weeks postinfection. A depressed response to concanavalin A (ConA) was seen at 1 week postinfection through week 5. Mitomycin C-treated cells from infected jirds were capable of suppressing the response of normal cells to ConA. Sephadex G-10-nonadherent spleen cells from infected jirds showed elevated responses to D. viteae antigen at 1, 3, and 5 weeks and elevated responses to ConA at 3 and 5 weeks. Filaria-specific antibodies were seen at 1 week postinfection, and titers rose through week 5. Plaque-forming cell production to sheep erythrocytes was not depressed in infected jirds. It was concluded that jirds react immunologically with both cellular and humoral responses during the prepatent period of D. viteae infection. A concurrent immune depression was seen. Its effect on resistance and tolerance remains to be determined.

Animals↗

Effect of response to backtest and housing condition on cell-mediated and humoral immunity in adult pigs.

Several recent studies in juvenile pigs demonstrated a relationship between the degree of resistance displayed early in life in a so-called "backtest" and parameters of cell-mediated and humoral immunity. Some of the immune characteristics were reported to depend on the interaction between backtest classification and housing system. In the present study, the effects of backtest classification and housing condition on immune reactivity in adult gilts were examined. At 10 and 17 days of age, female piglets were subjected to the backtest. In this test, each piglet is restrained on its back for 1 min and the number of escape attempts is scored. Pigs classified as high resisting (HR) or low resisting (LR) were selected and housed in groups of six gilts. At 7 months of age, half of the gilts were housed in individual stalls. At 12 months of age, gilts were challenged by immunization with DNP-KLH. Control gilts were treated similarly with a placebo. Blood samples were drawn prior to immunization (Day 0) and weekly thereafter until Day 28. No significant effects of backtest type on cellular and humoral responses against KLH were found. Furthermore, being housed in stalls as compared to groups had no consequences for the immune response and did not induce differences between HR and LR gilts. Differences in behavior and physiology found previously between HR and LR gilts, particularly in gilts in stall housing, may thus be of relatively little importance for immune-related health.

Adaptation, Physiological↗

A subcutaneously injected UV-inactivated SARS coronavirus vaccine elicits systemic humoral immunity in mice.

The recent emergence of severe acute respiratory syndrome (SARS) was caused by a novel coronavirus, SARS-CoV. It spread rapidly to many countries and developing a SARS vaccine is now urgently required. In order to study the immunogenicity of UV-inactivated purified SARS-CoV virion as a vaccine candidate, we subcutaneously immunized mice with UV-inactivated SARS-CoV with or without an adjuvant. We chose aluminum hydroxide gel (alum) as an adjuvant, because of its long safety history for human use. We observed that the UV-inactivated SARS-CoV virion elicited a high level of humoral immunity, resulting in the generation of long-term antibody secreting and memory B cells. With the addition of alum to the vaccine formula, serum IgG production was augmented and reached a level similar to that found in hyper-immunized mice, though it was still insufficient to elicit serum IgA antibodies. Notably, the SARS-CoV virion itself was able to induce long-term antibody production even without an adjuvant. Anti-SARS-CoV antibodies elicited in mice recognized both the spike and nucleocapsid proteins of the virus and were able to neutralize the virus. Furthermore, the UV-inactivated virion induced regional lymph node T-cell proliferation and significant levels of cytokine production (IL-2, IL-4, IL-5, IFN-gamma and TNF-alpha) upon restimulation with inactivated SARS-CoV virion in vitro. Thus, a whole killed virion could serve as a candidate antigen for a SARS vaccine to elicit both humoral and cellular immunity.

Adjuvants, Immunologic↗

Effect of androgens on the ontogeny of humoral immunity in chickens.

Steroids have significant influences on the immune system of birds and mammals, and the Food and Drug Administration should consider them in the regulatory review process. As discussed here, androgens have different effects depending on dose and the timing of administration. If given in sufficient amount during the critical time of bursal development, androgens may delay or permanently impair development of humoral immunity. If given after the bursa has developed, androgens apparently cause precocious migration without precocious maturation. Possibly, the cells fail to mature because of the absence of additional signals such as the immune modulating peptide hormones elaborated by reticuloepithelial cells of thymus and bursa. Even though the FDA requires that drugs for use in domesticated animals be tested in the species for which approval is being requested, chickens provide us with an inexpensive and useful experimental animal for preliminary studies of a drug's effect on the ontogeny of humoral and cellular immunity.

Animals↗

Cellular and humoral immunity in guinea pigs to two major polypeptides derived from hepatitis B surface antigen.

Guinea pigs immunized with hepatitis B surface antigen (HBsAg), types adw, adr and ayw, and with two major polypeptides derived from HBsAg/adw developed cell-mediated immunity as determined by the macrophage migration inhibition assay. Peritoneal exudate cells from animals immunized with a 22000- or a 25000-mol. wt. polypeptide derived from HBsAg/adw showed significant migration inhibition after challenge with either polypeptide or with purified HBsAg. Significant inhibition of macrophage migration was not observed when polypeptide-sensitized cells were challenged with normal human serum or with normal human liver extract. Similarly, a cell-mediated immune response was not observed in peritoneal exudate cells from animals sensitized to normal human serum or normal human liver extract which were challenged with either of the polypeptides. The humoral immune response to either of the polypeptides, as measured by radioimmunoassay, was substantially lower than that observed in animals immunized with intact particles. This apparent difference between cellular and humoral responses suggests that the macrophage migration assay is a sensitive indicator of the immunogenicity of the smaller mol. wt. HBsAg-derived polypeptides in guinea pigs.

Animals↗

The efficiency of humoral immune transfer depends on both the graft and the immunosuppressive treatment.

The present study was designed to compare the efficiency of adoptive transfer of humoral immunity after liver, kidney, and heart transplantation in relation to the number of passenger lymphocytes, and to estimate the risk of a detrimental effect and the chance of a beneficial effect. Hepatitis B virus surface-antigen-vaccinated brown Norway rats (BNs) and AxC 9935 Irish (ACI rats) served as donors, and naïve Lewis (LEW) rats as recipients. The liver grafts contained 100 times more passenger lymphocytes than heart grafts, and the kidney grafts approximately ten times more, indicated by monoclonal CD45 antibody staining. Transient anti-HBs immunity did occur after transplantation of all three organ grafts. In all rejecting groups, the serum recipient-to-donor anti-HBs titer ratio (R/D ratio) was below 0.10%, with heart recipients showing half the level (0.05%) of liver recipients (0.09%). Under immunosuppression, R/D ratio doubled in liver or kidney recipients, but remained unaffected in heart recipients. Immune transfer was most efficient in immune-suppressed liver recipients in the spontaneously tolerant strain combination as indicated by a significantly higher R/D ratio (0.32%) and a longer titer persistence (up to 9 weeks) than in all other groups. Therefore, mainly liver and kidney graft recipients carry a risk, but also a chance of benefiting from the transfer of donor-derived immunity.

Animals↗

Humoral immune and clinical responses to food antigens following acute diarrhoea in children.

OBJECTIVE: To investigate the effect of acute watery diarrhoea in children upon humoral immune responses to food antigens and the subsequent development of food allergy. METHODOLOGY: Serum antibodies to cows' milk, beta-lactoglobulin, alpha-lactalbumin, bovine serum albumin and ovalbumin were measured in 30 children with acute diarrhoea in the acute phase and 1 month after recovery. The children were followed for 1 year to assess the development of food allergy. RESULTS: IgG antibeta-lactoglobulin titres for the study group increased 1 month after recovery compared to the titres during the acute phase (P = 0.02). Antibody concentration for the other antigens studied did not rise. Four children developed positive IgE antibodies to one or more of the allergens after the diarrhoeal episode, although the titres were very low. None showed evidence of allergy to cows' milk or egg during the year-long follow-up. CONCLUSIONS: Acute diarrhoea in children resulted in increased production of IgG antibody to beta-lactoglobulin and had a priming effect for development of positive IgE antibody to cows' milk. Clinical food allergy was not observed in any of the children during the year-long follow-up.

Animals↗

Augmentation of in vitro humoral immune responses in the mouse by an antibody to IgD.

Heterologous anti-delta-chain antibodies have an adjuvant effect on specific in vivo humoral immune responses to simultaneously, or subsequently, injected antigens in the rat and rhesus monkey. We have used a hybridoma-secreted antibody that binds murine delta-chain of the allotype (4.22aM delta a) to study this phenomenon in the mouse and to investigate the mechanism of this effect. Injection of 4.22aM delta a into BALB/c mice removes almost all surface IgD (sIgD) from splenic B lymphocites. sIgD does not reappear until the serum level of 4.22aM delta a decreased 5-7 d after injection. 4.22aM delta a fails to induce detectable proliferation or to raise total serum Ig levels substantially above control values. However, 4.22aM dalta a injected 24 h before antigen elicits an approximately twofold enhancement of serum IgM and a 3- to 10-fold enhancement of serum IgG anti-trintriphenyl (TNP) antibodies in response to immunization with optimal doses of TNP-Ficoll or TNP-sheep red blood cells (TNP-SRBC). 4.22aM delta a injected 1 wk before or 3 d after TNP-SRBC, however, has no effect on IgG anti-TNP levels. The adjuvant effect of anti-delta-chain antibody was markedly decreased when suboptimal antigen doses were used. Furthermore, even in the case of TNP-Ficoll, a relatively T-independent antigen, the ability of 4.22aM dalta a to enhance the anti-TNP antibody response was T cell dependent. Our data suggest that the binding of anti-delta-chain antibody to cell sIgD may partially activate B lymphocytes and make them more capable of differentiating into antibody-secreting cells when stimulated by antigen-specific T cell help.

Animals↗

The humoral immune response and the productivity of laying hens kept on the ground or in cages.

The effects of two different keeping systems on the humoral immune response and productivity were compared for 80 laying hens, divided into four groups. Two groups each of 20 hens were kept on the ground and two were kept in cages. All the birds were immunised subcutaneously with human serum immunoglobulin G (IgG) at a dose of 100(microg per injection. The immunisations were performed twice at 4-week intervals. The lipopeptide Pam(3)Cys-Ser-(Lys)(4) was used as an adjuvant at a dose of 0.25mg per injection in one group from each housing system. In the second group from each housing system, the hens were immunised without any adjuvant (antigen control groups). The mean egg yield was significantly higher in both the antigen control group and the adjuvant group, when laying hens were kept in cages. Total egg weight remained constant in both of the housing systems. Keeping hens in cages resulted in higher mean specific antibody titres and mean immunoglobulin Y concentrations in the egg yolk.

Adjuvants, Immunologic↗

Humoral immune responses in Cr2-/- mice: enhanced affinity maturation but impaired antibody persistence.

Deficiency in CD21/CD35 by disruption of the Cr2 loci leads to impaired humoral immune responses. In this study, we detail the role of CD21/CD35 on Ab responses to the hapten (4-hydroxy-3-nitrophenyl)acetyl conjugated to chicken gamma-globulin. Surprisingly, Cr2-/- mice generate significant Ab responses and germinal center (GC) reactions to low doses of this Ag in alum, although the magnitude of their responses is much reduced in comparison with those of Cr2+/- and C57BL/6 controls. Increasing Ag dose partially corrected this deficit. In situ study of the somatic genetics of GC B cells demonstrated that VDJ hypermutation does not require CD21/CD35, and Cr2-/- mice exhibited enhanced affinity maturation of serum Ab in the post-GC phase of the primary response. On the other hand, Cr2-/- mice displayed accelerated loss of serum Ab and long-lived Ab-forming cells. These observations suggest that B cell activation/survival signals mediated by CD21 and/or the retention of Ag by CD21/CD35 play important roles in the generation, quality, and maintenance of serum Ab.

Amino Acid Sequence↗

[Humoral immune response in mice to hybrid nucleic acid vaccines containing Plasmodium falciparum merozoite surface protein 1 block 17-based gene].

AIM: To analyse the humoral immune response in mice to nucleic acid vaccines (VR1012/HG-MSP1-17 for intracellular expression or VR1012/TPA/HG-MSP-17 for secretion) containing Plasmodium falciparum merozoite surface protein 1 (MSP1) 17 block gene and gene fragment of several T cell epitopes from MSA1, MSA2, RESA, IL-1 and TT. METHODS: BALB/c or C57BL/6 mice received three times intramuscular immunization with 200 micrograms/100 microliters or 100 micrograms/100 microliters of VR1012/HG-MSP1-17 or VR1012/TPA/HG-MSP1-17 per mouse each time. Anti-HG or anti-MSP1-17 antibodies were monitored by indirect ELISA. RESULTS: BALB/c and C57BL/6 mice immunized with 100 micrograms/100 microliters of VR1012/HG-MSP1-17 per mouse raised significantly anti-HG and anti-MSP1-17 antibodies, but the levels of antibodies were not high. BALB/c mice immunized with 200 micrograms/100 microliters of VR1012/HG-MSP1-17 per mouse raised higher anti-HG antibodies but not anti-MSP1-17 antibodies. BALB/c mice immunized with 200 micrograms/100 microliters of VR1012/TPA/HG-MSP1-17 per mouse raised low level of anti-HG antibodies only. CONCLUSION: VR1012/HG-MSP1-17 is more immunogenic than VR1012/TPA/HG-MSP1-17.

Animals↗