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A family study of leprosy: subcutaneous amyloid deposits and humoral immune responses.

One group of 11 aboriginal families, consisting of 27 persons with leprosy and 43 unaffected family members, and a second group of 26 patients with leprosy were studied in the Northern Territory of Australia. Amyloid deposits were sought in fine needle aspirates of subcutaneous fat and serological investigations relevant to amyloidosis and to the humoral immune response were done. The study showed unexpectedly high frequencies of amyloid deposits, evidence of persisting hepatitis B virus (HBV) infection, and antibodies to smooth muscle, to skin basement membrane, and to extractable nuclear antigens (ENA). Compared with unaffected family members, patients with leprosy had increased serum alpha-lipoprotein (alpha-LP) and were more often hepatitis B surface antigen (HBsAg) carriers but, contrary to expectations, the presence of amyloid, the alpha-LP level, serum amyloid associated (SAA) protein, and the HBsAg carrier state all appeared unrelated to the type of leprosy.

Adult↗

Clinical outcome of lymphoma patients after idiotype vaccination is correlated with humoral immune response and immunoglobulin G Fc receptor genotype.

PURPOSE: The unique immunoglobulin idiotype (Id) expressed by each B-cell lymphoma is a target for immunotherapy. Vaccination with Id induces humoral and/or cellular anti-Id immune responses. However, the clinical impact of these anti-Id immune responses is unknown. We and others have previously reported that immunoglobulin G Fc receptor (FcgammaR) polymorphisms predict the clinical response of lymphoma patients to passive anti-CD20 antibody infusions. In this study, we tested whether anti-Id immune responses or FcgammaR polymorphisms associate with clinical outcome of patients who received Id vaccination. PATIENTS AND METHODS: We analyzed 136 patients with follicular lymphoma who had received Id vaccination. The anti-Id immune responses were measured and FcgammaRIIIa and FcgammaRIIa polymorphisms were determined and correlated with clinical outcome for these patients. RESULTS: Patients who mounted humoral immune responses had a longer progression-free survival (PFS) than those who did not (8.21 v 3.38 years; P = .018). Patients with FcgammaRIIIa 158 valine/valine (V/V) genotype also had a longer PFS than those with valine/phenylalanine (V/F) or phenylalanine/phenylalanine (F/F) genotypes (V/V, 8.21 v V/F, 3.38 years; P = .004; v F/F, 4.47 years; P = .035). Multivariate analysis using the Cox proportional hazards model showed that V/V genotype and humoral immune responses were independent positive predictors for PFS. CONCLUSION: This study is the first to identify the predictive value of FcgammaR polymorphism on clinical outcome in patients who received active immunotherapy with tumor antigen vaccines. Our results imply that the antibodies induced against a tumor antigen are beneficial and that FcgammaR-bearing cells mediate an antitumor effect by killing antibody-coated tumor cells.

Antigens, CD↗

Idiotypes of anti-Ia antibodies. I. Expression of the 14-4-4S idiotype in humoral immune responses.

The idiotype of a mouse monoclonal anti-I-E antibody, 14-4-4S, has been studied using a heterologous anti-idiotypic reagent. This antibody recognizes Ia. 7, an antigenic specificity present in all strains expressing a product of the I-E subregion. Expression of the 14-4-4S idiotype in humoral immune responses was analyzed by an idiotype-specific enzyme-linked immunosorbent assay system. The idiotype was readily detectable in C3H.SW anti-C3H alloantisera, the same immunization combination from which the hybridoma was derived. Absorption analysis demonstrated the anti-I-E specificity of the idiotype-positive molecules in these alloantisera. Penetrance of idiotype expression was high among individual C3H.SW immune mice (9 of 10 tested). To examine genetic requirements for idiotype expression, an immunization was performed using as responders CWB mice, congenic with C3H.SW but differing at the heavy chain allotype loci. Immune sera of individual CWB mice contained very little or no idiotype, demonstrating that levels of idiotype expression are influenced by allotype-linked genes, although the influence of other genes has not been ruled. The 14-4-4S idiotype therefore represents a shared idiotype of anti-Ia antibodies and provides opportunities for analysis of the idiotypes of cellular receptors for the corresponding Ia antigen.

Animals↗

Summary of antibody workshop: The Role of Humoral Immunity in the Treatment and Prevention of Emerging and Extant Infectious Diseases.

In the era before antibiotics, human diseases were commonly treated with immune animal and human sera, often with life-saving results. With the advent of emerging infectious diseases, many of which cannot be adequately treated or prevented, attempts to develop antibody treatments have taken on new importance. The role of humoral immunity in treatment and prevention was the focus of discussion at a 1996 workshop. The cellular and molecular mechanisms of neutralization were examined in detail. It was noted that success in passive immunity has frequently been the key element in devising a successful strategy to develop a vaccine for active immunization. The workshop concluded on a cautious note of optimism that antibody-based treatment and prevention for diseases such as human immunodeficiency virus infection, Ebola fever, and others of clinical and public health importance deserve further development and clinical trial.

Animals↗

Impairment of the thymus-dependent humoral immune response by syngeneic or allogeneic pregnancy.

Pregnancy results in a diminished capacity of the gravid mouse to respond to thymus-dependent antigens. The IgM response to thymus dependent and independent antigens is relatively unaltered whereas the IgG response to thymus-dependent antigens is markedly suppressed in both syngeneic and allogeneic pregnancy. The impairment of the humoral immune response is not due to loss of antibody-forming precursor lymphocytes (AFPL) from the spleen since their number remains unaltered. During pregnancy the spleen becomes enlarged and the precursor frequency is diluted somewhat but the AFPLs retain their normal activity in vitro. This implies an alteration in their amplification by accessory cells (T cells or macrophages) is responsible for the impaired IgG responses.

Animals↗

Identification of CLUAP1 as a human osteosarcoma tumor-associated antigen recognized by the humoral immune system.

Since the prognosis of human osteosarcoma in advanced stage remains poor, the development of new and effective therapies including immunotherapy is required. To identify tumor-associated antigens of osteosarcoma applicable to the immunotherapy of this malignancy, we employed the serological analysis of recombinant cDNA expression library (SEREX) technique that defines tumor antigens recognized by the humoral immune system. Screening a cDNA library derived from an osteosarcoma cell line MG63 with sera from osteosarcoma patients identified 43 positive clones, representing 14 distinct antigens. Among them, CLUAP1 (clusterin-associated protein 1) was highly expressed in osteosarcoma tissue samples and cell lines. Overexpression of CLUAP1 was observed in other malignancies including ovarian, colon, and lung cancers. Our results suggest that CLUAP1 may be useful as a prognostic/diagnostic marker and/or for a target of immunotherapy of osteosarcoma.

Adolescent↗

Modulation of the cellular and humoral immune responses of tumor patients by mistletoe therapy.

There is evidence from recent data that mistletoe extracts exert immunostimulatory properties which could explain their therapeutic effects observed in some tumor patients. Aim of our study was, therefore, to investigate the effect of a subcutaneous 16-weeks therapy with a mistletoe extract (ABNOBAviscum Mali, AM) on the cellular and humoral immune responses in eight breast cancer patients. Mistletoe therapy induced a strong initial proliferation of peripheral blood mononuclear cells (PBMC) in all individuals, which, however, decreased in six patients during the observation period, indicating that not only activating but also inhibitory mechanisms have been induced. In all supernatants of AM-stimulated cell cultures TNF-alpha or IL-6 were found, indicating the activation of cells of the monocyte-/macrophage lineage by mistletoe extracts. Further analyses revealed, that AM induced in vitro also the release of low amounts of IFN-gamma and IL-4 with individual variations. At the end of the therapy, a shift to Th1- related cytokines could be observed in the in vitro cell culture system. All patients produced anti-mistletoe lectin 1 antibodies of the IgG-type during therapy and in four of them additionally antibodies of the IgE-type were found. It, therefore, seems that AM can influence the Th1/Th2 balance and, in case of a Th1 shift, this may favourably influence the tumor growth.

Adjuvants, Immunologic↗

Neuroimmunomodulation of in vivo anti-rotavirus humoral immune response.

Neuropeptides and neurohormones (neurotransmitters) have been shown to modulate immune responses in vitro and in vivo. Since reproduction and lactation are regulated by neurohormones, we investigated whether neurohormones could enhance anti-rotavirus immunity in milk. Rotavirus-free mice were immunized orally with killed bovine rotavirus (BRV) and bred 6 weeks post-immunization. Post-whelping, each group of dams (ten mice/group) was given a single injection of prolactin (PRL), estrogen, PRL and estrogen or testosterone. The effects of neuropeptides, substance P (SP) and somatostatin (SS) on serum and lactogenic anti-rotavirus humoral immune responses were also investigated. The results revealed that in the groups given PRL or estrogen, anti-rotavirus antibody titers in milk and serum were enhanced. In contrast, testosterone had a negative effect on antibody titers. The administration of neuropeptide SP resulted in some enhancement of the lactogenic anti-rotavirus antibody titer at day 9 post-whelping whereas the opposite effect was observed following administration of SS. Prolactin given at 100 micrograms/mouse, on the day after whelping, gave optimum milk and serum antibody responses. Neurotransmitters potentiated immune responses to the weaker immunogenic proteins, VP4 and VP7 as well as to the strongly immunogenic VP6. In order to verify that the enhancement of anti-rotavirus antibody production was due to PRL and not to other factor(s), bromocriptine (BCR), a selective PRL inhibitor, was used as a control. Mice given BCR exhibited a drastic reduction in anti-rotavirus antibody in serum and milk. The role of neurotransmitters in the modulation of the lactogenic immune response and its significance in protection of neonates from enteric infections is discussed.

Animals↗

Immunoblot analysis of humoral immune response to Helicobacter pylori in children with and without duodenal ulcer.

Several studies have demonstrated that enzyme-linked immunosorbent assay is not a sensitive and specific method to diagnose Helicobacter pylori infection in children, especially in the younger ones. Since serum immune response can also be determined by immunoblotting and it permits the detection of antibodies to virulence factors such as CagA and VacA, we evaluated the accuracy of a commercial immunoblotting test to diagnose H. pylori infection and to assess the humoral immune response to different H. pylori antigens in 122 children who underwent upper gastrointestinal endoscopy. The presence of H. pylori was determined in antral biopsy specimens by culture, preformed urease test, and histological analysis. H. pylori was identified by microbiological and histopathological methods in 66 children (including all of the 21 who had duodenal ulcer). Antibodies to H. pylori were detected in 63 infected children and in 8 noninfected ones. The sensitivity, specificity, and positive and negative predictive values of the immunoblotting test were 95.5, 85.7, 88.7, and 94.1%, respectively. The number of immunoreactive bands increased with age (P = 0.003), and the bands of 35 kDa (P = 0.013); 89 kDa, the VacA antigen (P = 0.001); and 116 kDa, the CagA antigen (P = 0.00004) were more frequently observed in older children. The frequency of the bands of 89 kDa (P = 0.001) and 116 kDa (P = 0.03) was higher in children with duodenal ulcer than in H. pylori-positive children without the disease. In conclusion, the immunoblotting test appears to be useful for the diagnosis of H. pylori infection in children, even in the younger ones.

Adolescent↗

[The effect of experimental craniocerebral trauma on cell-mediated and humoral immunity reactions].

Brain injury (BI) was induced in 9 rabbits either after cranial trepanation or through graded blow. Cell-mediated immunity to brain autoantigens in the automix-culture test and the production of specific antibodies on soluble protein antigen immunization were studied. Experiment BI was not found to result in the emergence of the lymphocytes capable of mediate cellular autoimmune responses. However, there were increases in the nonspecific lymphocytic sensitivity to the T-lymphocytic mitogen ConA and in a humoral immune response, suggesting the activation of T- and B-cell immunity following BI.

Animals↗

Association of some HLA-DRB1 antigens with Echinococcus granulosus specific humoral immune response.

An ELISA system using crude camel hydatid fluid antigen was used to detect specific IgG and IgG1 in sera of 35 cystic echinococcosis (CE) patients in whom the distribution of class II HLA-DR3 and HLA-DR11 was determined. The recorded sensitivities were 88.6 % and 94.35 % for IgG and IgG1 respectively. In patients with high humoral immune response a statistically highly significant increased frequency of HLA-DR3 (P<0.01) was recorded for IgG with high relative risk value (RR=3.2) and reasonable etiologic fraction (EF=0.3), while HLA-DR11 recorded P<0.05, RR=2.6, & EF=0.2. For IgG1 both antigens showed significant increased frequency (P<0.05), RR (2.95 & 2.79 respectively), EF (0.28 & 0.23 respectively). HLA-DR3 was highly significantly associated with complicated cases (P<0.01, RR=4.36, EF=0.4) in whom the mean antibody units for both IgG and IgG1 were significantly raised (P<0.05). It is advisable to rely on IgG1 for the diagnosis of CE and to consider the genetic disposition of the patient as an important criterion in the outcome of infection.

Animals↗

[Characteristics of heterologous membrane antigen (HM Ag) and humoral immune response to HM Ag in pregnant women and patients with gynecologic malignancies].

Heterologous membrane antigen (HM Ag) is discovered to be incorporated into membranes of cultured human cells grown in media containing fetal calf serum (FCS). To know the characteristics of HM Ag, we investigated the dynamics of HM Ag on cell membrane, distribution of HM Ag and humoral immune response to HM Ag in pregnancy and gynecologic malignancies. The following results were obtained. HM Ag disappeared from the cell membranes of M14 grown in media with fetal calf serum (FCS) after the cells were transferred into whole human serum (12 days) or into a gamma human serum (18 days). M14 cells grown in human serum reincoporated HM Ag into their membranes within 48 hours after transfer to medium with 20% FCS. HM Ag was detected in FCS, adult bovine serum, cow's milk and erythrocytes homogenate of ox and sheep. The natural anti-HM Ag antibody appeared frequently in pregnant women, patients with gynecologic malignancies and healthy persons. These findings suggest that HM Ag must be excluded in immunologic studies of cultured human cancer cells by changing the medium from FCS to human serum or by absorbing anti-HM Ag antibody with cells carrying HM Ag.

Antigens, Surface↗

Humoral immune response against Fasciola gigantica fatty acid binding protein.

Recombinant fatty acid binding protein of Fasciola gigantica was expressed in Escherichia coli and purified by nickel chelating affinity chromatography. The recombinant protein along with native fatty acid binding protein (FABP) isolated from the parasite were evaluated for their potential in the diagnosis of F. gigantica infection in sheep, cattle and buffaloes, both by ELISA and western blotting. Results of this study indicate that there is no humoral immune response generated against this protein in the experimental infection of these ruminants with F. gigantica, thereby limiting the usefulness of this antigen in the early diagnosis of fasciolosis in these animals. Also, the paper discusses the probable reasons for the failure of this protein in detecting humoral response in these animals by ELISA and immunoblotting.

Animals↗

A graph model for the evolution of specificity in humoral immunity.

The immune system protects the body against health-threatening entities, known as antigens, through very complex interactions involving the antigens and the system's own entities. One remarkable feature resulting from such interactions is the immune system's ability to improve its capability to fight antigens commonly found in the individual's environment. This adaptation process is called the evolution of specificity. In this paper, we introduce a new mathematical model for the evolution of specificity in humoral immunity, based on Jerne's functional, or idiotypic, network. The evolution of specificity is modeled as the dynamic updating of connection weights in a dynamic graph whose nodes are related to the network's idiotypes. At the core of this weight-updating mechanism are the increase in specificity caused by clonal selection and the decrease in specificity due to the insertion of uncorrelated idiotypes by the bone marrow. As we demonstrate through numerous computer experiments, for appropriate choices of parameters the new model correctly reproduces, in qualitative terms, several immune functions.

Antibody Formation↗

Genetic variations and humoral immune responses to myelin oligodendroglia glycoprotein in adult phenotypes of X-linked adrenoleukodystrophy.

The lack of phenotype/genotype association in X-linked adrenoleukodystrophy (X-ALD) has prompted the search for disease modifying factors. We previously demonstrated increased serum antibody responses against myelin oligodendrocyte glycoprotein (MOG) in various clinical phenotypes of X-ALD allowing speculations that myelin specific humoral immune responses might be involved in phenotype generation of X-ALD. In the present study, we investigated the possible association of (1) a naturally occurring variable number tandem repeat (vntr) polymorphism (C allele) in the 3' flanking region of the interleukin-6 gene (IL-6), previously demonstrated to modify the course of Alzheimer's disease, systemic lupus erythematodes and Multiple Sclerosis (MS), (2) a tetranucleotide repeat polymorphism (TAAA)(n) in the 3' flanking region of the MOG gene and (3) HLA class II alleles with adult clinical phenotypes and serum antibody responses to MOG in 70 adult X-ALD patients. HLA class II alleles, (TAAA)(n) MOG gene polymorphisms, and IL-6 C allele were not associated with clinical phenotypes. Anti-MOG antibodies were detectable in 17/54 X-ALD patients (31.5%). Anti-MOG antibodies were associated with the 226 bp (TAAA)(n) MOG gene polymorphism but not with distinct clinical phenotypes.

Adrenoleukodystrophy↗

Antigenic differences between metastatic cells in bone marrow and primary tumours and the anti-MUC1 humoral immune response induced in breast cancer patients.

The dissemination of a malignant neoplasia is a complex process, which requires a set of molecules that remains unknown. It has been suggested that mucins and their carbohydrate-associated antigens may be implicated in tumour spreading which may be also influenced by an anti-MUC1 immune response. In this pilot study, we report the pattern of carbohydrate and peptidic MUC1-associated epitopes on carcinoma cells isolated from bone marrow (BM), taking into account primary tumour histopathologic features. We also bring information about the anti-MUC1 humoral response in these patients. Seventeen patients with invasive breast carcinoma were included. A sample of the primary tumour, a serum sample and a BM aspirate were obtained from each patient. Clinical features studied were tumour size, number of metastatic nodes, histological type and disease stage. Standard immunohistochemistry was performed with antigenic retrieval using different monoclonal antibodies (MAbs): anti carbohydrate antigens: Lewis x (KM380), sLewis x (KM93), Lewis y (C14) and Tn, anti-MUC1 peptide core MAbs: C595, HMFG2 and SM3, anti-cytokeratins, anti-protoncogenes ErbB2 and ErbB3 (IgG) MAbs and also anti-CD34 and anti-CD45 MAbs. ELISA techniques were employed to study circulating MUC1 as well as free and complexed anti-MUC1 antibodies. Immunohistochemical results showed that carbohydrate antigenic expression increases in BM neoplastic cells compared to the original tumours. However, we were not able to demonstrate that a humoral immune response to MUC1 has been induced in these patients. Finally, the employed procedures allow the selective immortalisation of micrometastatic carcinoma cells since short-term cell lines were established.

Antibodies, Monoclonal↗

Cross-protection of Salmonella abortusovis, S. choleraesuis, S. dublin and S. gallinarum in mice induced by S. abortusovis and S. gallinarum: bacteriology and humoral immune response.

Cross-protection induced by primary infection with Abortusovis and Gallinarum was examined against challenge injection with these Salmonella serotypes as well as with Dublin and Choleraesuis, the other virulent serotypes. Abortusovis induced efficient protection against the other Salmonella. Gallinarum was ineffective against Choleraesuis. Even with low multiplication in mice, the Gallinarum J91 strain induced a weak but significant protection against Dublin (same O group serotype). The antibodies in the blood of mice were tested with ELISA specific for the Salmonella antigens used to prime or to challenge animals. The Gallinarum J91 strain was detected to be more antigenic in ELISA than the other Salmonella antigens. It is difficult to conclude on a correlation between IgM or IgG antibodies and induction of protection, because of the variability in immune response according to the different serotype used. Nevertheless, the negative linkage between a number of bacteria in the spleen of mice challenged with Gallinarum and Dublin, and the level of IgM and IgG antibodies specific for the challenging serotype, showed that humoral immune response could be one element of cross-protection, mainly by the immune response against the same O serotype.

Animals↗

Humoral immunity to adeno-associated virus type 2 vectors following administration to murine and nonhuman primate muscle.

Adeno-associated virus (AAV) is being developed as a vector capable of conferring long-term gene expression, which is useful in the treatment of chronic diseases. In most therapeutic applications, it is necessary to readminister the vector. This study characterizes the humoral immune response to AAV capsid proteins following intramuscular injection and its impact on vector readministration. Studies of mice and rhesus monkeys demonstrated the formation of neutralizing antibodies to AAV capsid proteins that persisted for over 1 year and then diminished, but this did not prevent the efficacy of vector readministration. More-detailed studies strongly suggested that the B-cell response was T cell dependent. This was further evaluated with a blocking antibody to human CD4, primatized for clinical trials, in a biologically compatible mouse in which the endogenous murine CD4 gene was functionally replaced with the human counterpart. Transient pharmacologic inhibition of CD4 T cells with CD4 antibody prevented an antivector response long after the effects of the CD4 antibody diminished; readministration of vector without diminution of gene expression was possible. Our studies suggest that truly durable transgene expression (i.e., prolonged genetic engraftment together with vector readministration) is possible with AAV in skeletal muscle, although it will be necessary to transiently inhibit CD4 T-cell function to avoid the activation of memory B cells.

Animals↗