Increased and prolonged production of specific polymeric IgA after systemic immunization with tetanus toxoid in IgA nephropathy.

IgA nephropathy (IgAN) is a chronic form of glomerulonephritis which is characterized by the deposition in the glomerular mesangium of polymeric IgA (pIgA), the source of which is unknown. In order to investigate the production of pIgA in IgAN, patients were immunized systemically with tetanus toxoid (TT). Two weeks after immunization patients and controls responded to TT with an IgA response of similar magnitude. HPLC separation of sera showed that patients with IgAN produce significantly more pIgA anti-TT than controls (7.7 versus 2.88 arbitrary units; P less than 0.04). At this time, 33% of serum IgA anti-TT produced by patients with IgAN was polymeric, compared with 21% produced by controls (P less than 0.02). Monomeric IgA (mIgA) anti-TT levels were similar in both groups. Four weeks after immunization the proportion of pIgA anti-TT in controls and patients was significantly reduced from the 2 week level (from 21% to 0%, P less than 0.02 for controls; and from 33% to 8%, P less than 0.001, for patients). Only four out of 12 controls had any detectable pIgA anti-TT at this time compared with nine out of 10 patients with IgAN (P less than 0.05), and IgAN patients produced proportionally more pIgA anti-TT than did controls (median 8%, interquartile ranges (IQR) 4-10% versus 0% IQR 0-3%; P less than 0.01). HPLC analysis under acid conditions did not alter the pattern of pIgA and mIgA anti-TT, suggesting that the high molecular weight IgA fraction was not due to complexes. These data indicate that circulating pIgA results (at least in part) from a systemic response to antigen, which may be exaggerated in IgAN.     

T and B cell responses following immunization with tetanus toxoid in IgA nephropathy.

The B and T cell responses were investigated in IgA nephropathy before and after immunization with tetanus toxoid (TT). Both IgA and IgG anti-tetanus toxoid antibodies were elicited, but the IgA antibodies were significantly greater in patients (92.6 +/- 11.7 ELISA units) than in the controls (49.2 +/- 7.5 ELISA units). This was associated with a significantly greater proportion of IgA+ B cells in patients than controls before immunization. However, a significant increase in the proportion of IgA1 binding CD4 and CD8 cells was also found. The proportion of CD3 cells with gamma delta T cell receptors (CD3+TCR gamma delta +), was significantly greater before immunization in the IgA nephropathy patients (37.0% +/- 2.4), compared with controls (10.0% +/- 2.3; P less than 0.001). Immunization with TT further enhanced the CD3+TCR gamma delta + cells in patients to 45.8% +/- 7.2 compared with controls (16.3% +/- 4.5), with a corresponding decrease in CD3+TCR alpha beta + cells in the patients (P less than 0.001). CD3+TCR gamma delta + cells are upregulated by common microbial antigens and clinical exacerbations of IgA nephropathy are frequently associated with mucosal infections and a rise in serum IgA concentration. The increased TCR gamma delta expression may be responsible for the enhanced IgA antibody response in IgA nephropathy. The increase in IgA antibodies may than exert a controlling effect by binding to augmenting T cells and thereby inhibiting their function.     

Increased dosage of diphtheria toxoid for basic immunization of adults

Basic immunization of adults with increased dosages of a diphtheria toxoid vaccine (2100 flocculation units (Lf)/mg) was evaluated. Three injections of 7.5 Lf or 15 Lf diphtheria toxoid were given to 243 adults who had a history of no more than one previous vaccine injection. Systemic reactions were rare in both groups. Following the first two injections, local reactions (>5 cm) were observed in 6–14% of the adults. After the third injection, 35% of adults in the 15 Lf group reported a local reaction (>5 cm) compared to 11% in the 7.5 Lf group (p<0.001). The 15 Lf dose elicited a better antitoxin response than the 7.5 Lf dose. In a seronegative subgroup including 65 vaccinees who showed no booster response to the first vaccination, 79% had a postvaccination titer of 0.1 IU/ml and 28% a titer of 1.0 IU/ml after the third injection of 7.5 Lf. The corresponding numbers in the 15 Lf group were 94% and 44%, respectively. The study demonstrates that 7.5 Lf and 15 Lf diphtheria toxoid of high purity can safely be given to adults for basic immunization. The higher dose is more immunogenic but local reactions increase after the third injection.     

The effect of passive immunization against tetanus toxoid in man.

The present study deals with the immunological response in man after passive immunization against tetanus toxoid. Treatment of man with equine antitetanus serum stimulates a rise of E rosette-forming T lymphocytes. The level of IgG also rises while the level of IgM falls. It is assumed that the foreign globulin acts as an antigen that evokes the T lymphocytes' co-operation for the humoral immune response with a switch from IgM to IgG synthesis.     

Appearance of IgG and IgA antibodies in human bile after tetanus toxoid immunization

Humans immunized intramuscularly with one dose of tetanus toxoid exhibited IgG, and in some cases IgA antibody, in their bile as well as serum. Both isotypes appeared in bile transiently with titres declining after about day 10 for both classes. These kinetics resembled those of the serum IgA response but were markedly different to those for IgG antibody in serum. Measured IgG titres in bile were between 0.07 and 4.2% of those in paired sera, and IgA titres were between 6.8 and 124% of sera. Peak responses in bile, while generally of smaller size, exceeded those of paired sera when expressed as antibody/mg of IgG or IgA present. This calculation showed that during the peak response bile was up to nineteen-fold more abundant in IgG antibody than was serum taken at the same time, and up to forty-five-fold more for IgA. Enrichment of antibody in bile is not consistent with the Ig of bile being solely conferred by plasma, and may mean the involvement of local synthesis too. This study indicates that tetanus toxoid immunization of humans results in biliary antibody and raises the possibility of intra-hepatic antibody production for export to the intestinal tract in man.     

Ex vivo monitoring of antigen-specific CD4+ T cells after recall immunization with tetanus toxoid

To monitor antigen-specific CD4+ T cells during a recall immune response to tetanus toxoid (TT), a sequential analysis including ex vivo phenotyping and cytokine flow cytometry, followed by cloning and T-cell-receptor (TCR) spectratyping of cytokine-positive CD4+ T cells, was performed. Grossly, twice as many TT-specific CD4+ T-cell clones, ex vivo derived from the CCR7+/- CD69+ interleukin-2-positive (IL-2+) CD4+ subsets, belonged to the central memory (T(CM); CD62L+ CD27+ CCR7+) compared to the effector memory population (T(EM); CD62L- CD27- CCR7-). After the boost, a predominant expansion of the T(CM) population was observed with more limited variations of the T(EM) population. TCR beta-chain-variable region (BV) spectratyping and sequencing confirmed a large concordance between most frequently expressed BV TCR-CDR3 from ex vivo-sorted CCR7+/- CD69+ IL-2+ CD4+ subsets and BV usage of in vitro-derived TT-specific CD4+ T-cell clones, further demonstrating the highly polyclonal but stable character of the specific recall response to TT. Taken together, ex vivo flow cytometry analysis focused on the CCR7+/- CD69+ IL-2+ CD4+ subsets appears to target the bulk of antigen-specific T cells and to reach an analytical power sufficient to adequately delineate in field trials the profile of the antigen-specific response to vaccine.     

Peripheral lymphocytes changes in the anamnestic response to tetanus toxoid challenge.

Few data are available on the blood lymphocyte response to revaccination in man. The anamnestic response to tetanus toxoid challenge was evaluated by a variety of techniques during the first week after revaccination. Out of twenty subjects used, eight were evaluated before and 5 days after the injections (days 1--8). Analytical cell electrophoresis showed important variation in the B and two T lymphocyte populations. The B cell percentages, assessed by EAC-rosettes and electrophoretic mobility, were found to decrease by days 2 and 3, and return to former levels by day 8, when a rise in specific antibodies was detected. A similar response was found in the T1 population generally considered to be composed of low affinity E-rosette-forming cells. Conversely, a significant increase (50--100%) in circulating T2 lymphocytes (active rosette-forming cells) was found by days 2 and 3, followed by a rapid decrease of these 'differenciated' cells. The increase in the T2 lymphocytes appeared earlier in skin test positive subjects. These changes were correlated with E-rosettes, mitogen stimulation, peripheral leucocyte migration inhibition and transformation in the presence of the antigen. EA-IgG rosettes and ADCC varied similarly. These results may indicate a significant non-specific cell mobilization following revaccination.     

Loss of circulating T lymphocytes with normal levels of B and ''null'' lymphocytes in Thai adults with malaria.

Peripheral blood mononuclear cells from forty-nine Thai adults infected with either Plasmodium falciparum or Plasmodium vivax were examined in order to determine the percentage of T, B, and Fc-receptor-bearing cells present. In comparison to healthy controls, both the percentage and concentration of peripheral T cells were decreased in the malaria-infected individuals as assessed by formation of rosettes with sheep red blood cells. The percentage of peripheral B cells was increased but their concentration was unchanged, as assessed by two techniques: the presence of surface immunoglobulin and the presence of a complement receptor. Both the percentage and concentration of lymphocytes bearing Fc receptors were unchanged in infected individuals. Finally, calculation of the changes in 'null' cells (defined either as non-T, non-B lymphocytes or as non-T, non-B, non Fc-receptor-bearing lymphocytes) revealed an increase in the 'null' cell percentage but a decrease in the absolute number of 'null' cells. These data indicate that in adult Thai patients naturally infected with malaria, there is a real loss of circulating T lymphocytes with no real change in B, Fc-receptor-bearing, or 'null' lymphocytes.     

The plasmocytic reaction and immunological laws communication II. Immunological inhibition during immunization of rabbits with tetanus toxoid

Summary This investigation was devoted to the study of the inhibitory process in rabbits caused by repeated administration of tetanus toxoid. Investigations demonstrated that cytological changes develop in the regional lymphatic nodes of rabbits on repeated injection of the toxoid in sensitized animals. These changes are characterized by the accumulation of large cells of lymphoid type with a basophilic protoplasm (lymphoblasts or plasmoblasts). The serological examination of the extracts of these nodes shows the accumulation of large amounts of the antitoxin even in the early periods.Numerous repeated revaccination with the toxoid causes a condition of immunological inhibition in rabbits, which is serologically manifested by the cessation of the rise of the antitoxin concentration in the regional nodes and later in its drop.Morphologically this inhibition is manifested in the absence of the characteristic increase of the lymphoblasts in the lymph nodes. The effect of revaccination was strictly local. The content of antitoxin in the regional lymphatic nodes was low irrespective of the number of revaccinations.Presented by Active Member of the AMN SSSR P. F. Zdrodovskii     

Simultaneous administration of diphtheria-tetanus-pertussis-polio and hepatitis B vaccines in a simplified immunization program: immune response to diphtheria toxoid, tetanus toxoid, pertussis, and hepatitis B surface antigen.

We studied the interactions of hepatitis B vaccine with other vaccines used in the World Health Organization expanded programs of immunization. Three groups of Senegalese children were vaccinated with hepatitis B vaccine (HB) alone, diphtheria-tetanus-pertussis (DTP)-polio vaccine alone, or a combination of hepatitis B vaccine and DTP-polio vaccines simultaneously. The immune responses to HBsAg, tetanus toxoid, diphtheria toxoid, and pertussis were measured after one and two vaccinations at 6-month intervals. The immune responses to the combination of HB vaccine and DTP-polio vaccines were similar to the immune responses observed after administration of each vaccine alone. In addition, no adverse reactions were noted. These experimental trials also demonstrated that with a DTP-polio vaccine containing 30Lf of tetanus and diphtheria toxoids, two doses given at 6-month intervals are sufficient to provide a satisfactory immune response. In the case of pertussis and HB vaccines; however, a third dose is necessary.     

Increased EA-rosette formation by lymphocytes from patients with rheumatoid arthritis.

A modified method for estimating erythrocyte-antibody (EA) rosette formation of human peripheral blood lymphocytes (PBL) reveals consistent differences between rheumatoid arthritis (RA) patients tested and healthy control subjects. Using this method we find an average of 27 +/- 0-8% (standard error of mean) of PBL from 120 RA patients forming EA rosettes in contrast to only 6 +/- 0-6% of PBL from ninety-five healthy controls, and 7 +/- 0-9% from eighteen patients with systemic lupus erythematosus (SLE). This difference is not due to monocytes forming EA rosettes or to T-cell sheep red blood cell (SRBC) binding. The concentration of antibody used in our assay appears to highlight the RA-control differences--suggesting a possible qualitative difference in EA-binding capacity. We find no correlation between EA binding and disease duration or rheumatoid factor titre. The assay is susceptible to technical variation, and the effects of antibody concentration, lymphocyte to SRBC ratio, method of blood collection and lymphocyte-separation procedure have all been evaluated.     

Increased production of antigen-specific B lymphocytes during in vitro immunization using carrier-specific T helper hybridomas.

An in vitro method to increase the production of hapten-specific antibody-forming B cells (AFC) using a carrier-specific T helper hybridoma and murine splenocytes is described. Naive splenocytes (6 x 10(6)/ml) are cultured in vitro in the presence of a hapten-carrier conjugate (DNP.OVA) and OVA-specific T helper hybridomas (0.5 x 10(6)/ml). After 4-5 days in vitro immunization (IVI), the maximum number of DNP-specific AFC were found using a spot-ELISA with twice the number of IgM positive cells as IgG positive AFC. The presence of antigen in the form of a hapten-carrier complex and the use of a carrier-specific Th hybridoma resulted in more hapten-specific AFC than when neither antigen nor Th hybridoma were present or when antigen alone or T help alone were used. Also when the hapten was conjugated to a carrier not recognised by the carrier-specific Th hybridoma there were considerably fewer (less than 50%) hapten specific AFC formed. When in vivo primed splenocytes (DNP) were boosted in vitro (IVB) under the same conditions as for IVI most hapten-specific AFC were found on day 4 and both anti-DNP IgM and IgG AFC were increased relative to IVI. Again most AFC were found when hapten was bound to the relevant carrier. In conclusion, carrier-specific T hybridomas can be used in an in vitro immunization procedure with naive or primed splenocytes to increase the frequency of anti-hapten AFC. This method offers an improvement over the current in vitro immunization procedures for the production of monoclonal antibodies.     

A vaccine against tetanus with a higher level of tetanus toxoid

In an immunological survey of the population it was revealed that the population above 60 years is not adequately protected against tetanus. This is probably to a considerable extent due to changes in the immune system as a result of age-conditioned changes and also due to an inadequate stimulus in the form of vaccine. The author prepared therefore an anti-tetanus vaccine with a higher antigen content. After its administration to a group of elderly subjects experimental evidence was provided of its favourable effect on the formation of tetanic antitoxin. The main attention was focused on the effect of vaccine in prophylaxis of tetanus after injuries.     

Impaired human responses to tetanus toxoid in vitamin A-deficient SCID mice reconstituted with human peripheral blood lymphocytes.

Vitamin A deficiency is associated with increased childhood morbidity and mortality from respiratory and diarrheal diseases. In order to evaluate the effect of vitamin A on human antibody responses, we developed a vitamin A-deficient severe combined immunodeficient (SCID) mouse model. Vitamin A-deficient mice were produced by depriving them of vitamin A at day 7 of gestation. Mice were reconstituted with human peripheral blood lymphocytes (huPBL) from tetanus toxoid immune donors at 6 weeks of age and immunized with tetanus toxoid at 6 and 8 weeks of age. Secondary human antibody responses were determined 10 days later. The geometric mean human anti-tetanus toxoid immunoglobulin G concentrations were 3.75 micrograms/ml for the deficient mice and 148 micrograms/ml for controls (P = 0.0005). Vitamin A-deficient mice had only a 2.9-fold increase in human anti-tetanus toxoid antibody compared with a 74-fold increase in controls (P < 0.01). Supplementation with vitamin A prior to reconstitution restored human antibody responses to normal. These data suggest that vitamin A deficiency impairs human antibody responses. We speculate that impaired responses could increase susceptibility to certain infections. Furthermore, we propose that effects of other nutritional deficiencies on the human immune system could be evaluated in the SCID-huPBL model.     

The stimulation of IgM rheumatoid factor from human B lymphocytes by rheumatoid arthritis complement-activating immune complexes.

Immune complexes from rheumatoid arthritis (RA IC) and Hodgkin's disease (HD IC) sera were separated on an anti-C3g affinity column and their ability to stimulate the production of IgM and IgM RF by normal and RA B lymphocytes tested in a culture system in vitro. RA IC stimulated IgM production of which up to 91.3% had IgM RF activity. HD IC were incapable of stimulating the production of IgM and IgM RF. The stimulation of IgM and IgM RF production by RA IC required de novo protein synthesis. Both RA IC and HD IC were capable of significantly inhibiting (from 47.6 to 72.0%) pokeweed mitogen (PWM)-induced and goat F(ab)2 antihuman mu-induced B lymphocyte proliferation. Thus it is proposed that IC present in human pathological sera may regulate immunoglobulin production by an effect on B lymphocyte proliferation while some may, in addition, be capable of inducing IgM RF production from such cells.     

Total and IgE antibody levels following booster immunization with aluminum absorbed and nonabsorbed tetanus toxoid in humans

Total and IgE serum antibodies to tetanus toxoid were measured in 32 healthy adults, 3-4 weeks following booster immunization with either plain or aluminum hydroxide-absorbed tetanus toxoid. Whereas no difference in the total antibody values was observed, the level of anti-tetanus toxoid IgE antibodies was significantly higher in the group boostered with the adjuvanted vaccine.