Humoral immune response of patients bitten by the snake Bothrops erythromelas / Resposta imune humoral em pacientes picados pela serpente Bothrops erythromelas

INTRODUCTION: Snake envenomings are a health problem in rural areas of tropical and subtropical countries, but little is known regarding the immune response presented by bitten individuals. The IgM production of patients bitten by Bothrops erythromelas snake was analyzed to identify the effectiveness of treatment in this type of envenomation. METHODS: Bothrops erythromelas venom was submitted to electrophoresis and transferred to a nitrocellulose sheet, following incubation with patients' sera. RESULTS: A 38 KDa protein was detected before and 24 h after therapy. CONCLUSIONS: The result suggests that this protein could be used as a marker for individuals envenomed by Bothrops. erythromelas.

INTRODUÇÃO: Envenenamentos ofídicos consistem problema de saúde pública em áreas rurais de países tropicais e subtropicais, mas pouco sabe-se sobre a resposta imune apresentada pelos indivíduos picados, por isso a avaliação da produção de IgM por pacientes picados por Bothrops erythromelas identificando a eficácia do tratamento nesse tipo de envenenamento. MÉTODOS: O veneno de Bothrops erythromelas foi submetido a eletroforese e transferido para nitrocelulose, seguindo incubação com soro de pacientes. RESULTADOS: Foi observada proteína de 38KDa antes e 24 horas após o tratamento. CONCLUSÕES: Os resultados sugerem que essa proteína poderia ser utilizada como marcador para indivíduos envenenados pela serpente Bothrops erythromelas.

Time gap between oocyst shedding and antibody responses in mice infected with Cryptosporidium parvum

We observed the time gap between oocyst shedding and antibody responses in mice (3-week-old C57BL/6J females) infected with Cryptosporidium parvum. Oocyst shedding was verified by modified acid-fast staining. The individually collected mouse sera were assessed for C. parvum IgM and IgG antibodies by enzyme-linked immunosorbent assay from 5 to 25 weeks after infection. The results showed that C. parvum oocysts were shed from day 5 to 51 post-infection (PI). The IgM antibody titers to C. parvum peaked at week 5 PI, whereas the IgG antibody titers achieved maximum levels at week 25 PI. The results revealed that IgM responses to C. parvum infection occurred during the early stage of infection and overlapped with the oocyst shedding period, whereas IgG responses occurred during the late stage and was not correlated with oocyst shedding. Hence, IgM antibody detection may prove helpful for the diagnosis of acute cryptosporidiosis, and IgG antibody detection may prove effective for the detection of past infection and endemicity.

Rat hippocampus and primary immune response.

During immune challenge hippocampal region shows time-dependent changes in neurotransmitter levels. Hence in the present study the effect of electrolytic lesion in the dorsolateral hippocampus (DLH) and ventral hippocampal formation (VHF) (to create a disturbance in neurotransmitter levels) on humoral immunity in albino rats has been studied along with appropriate controls. Haemagglutination titre, IgM and IgG levels were monitored on the 5th day after an immune challenge by sheep red blood cells (SRBC) suspension. Antigen challenged lesioned animals had low circulating antibody titre levels compared with the controls and their site-specific sham lesioned groups. The IgM levels were significantly lowered in both DLH and VHF lesioned and immunized animals compared to their immunized sham groups as well as immunized controls. However, only immunized VHF lesioned group showed a significant decrease in IgG level from their immunized sham group. It was concluded from the results that an intact hippocampal region is essential for the normal humoral immunity for the primary immune response in rats. Probably VHF region may be required for the secondary immune response as indicated by the alteration in IgG levels in these animals.

Isotype specific immune responses in murine experimental toxocariasis

In this work, a murine experimental model of toxocariasis has been developed in BALB/c, C57BL/10 and C3H murine strains orally inoculated with 4,000 Toxocara canis embryonated eggs, in order to investigate the isotype-specific immune responses against excretory-secretory antigens from larvae. T. canis specific IgG+M, IgM, IgG, IgA, IgG1, IgG2a and IgG3 were tested by ELISA. The dynamics of the specific immunoglobulins (IgG+IgM) production showed a contrasting profile regarding the murine strain. Conversely to the results obtained with the IgM isotype, the IgG antibody class showed similar patterns to those obtained with IgG+IgM antibodies, only in the case of the BALB/c strain, being different and much higher than the obtained with IgG+IgM antibodies, when the C3H murine strain was used. The antibodies IgG+IgM tested in BALB/c and C57BL/10 were both of the IgM and IgG isotypes. Conversely, in the C3H strain only IgG specific antibody levels were detected. The IgG1 subclass responses showed a similar profile in the three murine strains studied, with high values in BALB/c, as in the case of the IgG responses

Production of monoclonal antibodies to a tumor--associated antigen by spontaneous cell fusion.

Spontaneous cell fusion induced by the bacterium Haemophilus paragallinarum has been recently reported as an alternative technique to generate hybridomas producing monoclonal antibody (mAb). In order to investigate the advantages of this technique to produce anti-tumor monoclonal antibodies we performed comparative experiments between H. paragallinarum induced spontaneous cell fusion and polyethylene glycol (PEG) mediated fusion. Hybridomas producing monoclonal antibodies to an experimental murine lymphoma antigen, the Dalton's lymphoma associated antigen (DLAA) were generated and their sensitivity and specificity were ascertained. The spontaneous fusion yielded more number of stable and specific hybridomas than PEG mediated fusion. The results suggest the advantage of H. paragalinarum induced cell fusion for the simplified production of specific antitumor monoclonal antibodies.

Characterization of the mucosal and systemic immune response induced by Cry1Ac protein from Bacillus thuringiensis HD 73 in mice

The present paper describes important features of the immune response induced by the Cry1Ac protein from Bacillus thuringiensis in mice. The kinetics of induction of serum and mucosal antibodies showed an immediate production of anti-Cry1Ac IgM and IgG antibodies in serum after the first immunization with the protoxin by either the intraperitoneal or intragastric route. The antibody fraction in serum and intestinal fluids consisted mainly of IgG1. In addition, plasma cells producing anti-Cry1Ac IgG antibodies in Peyer's patches were observed using the solid-phase enzyme-linked immunospot (ELISPOT). Cry1Ac toxin administration induced a strong immune response in serum but in the small intestinal fluids only anti-Cry1Ac IgA antibodies were detected. The data obtained in the present study confirm that the Cry1Ac protoxin is a potent immunogen able to induce a specific immune response in the mucosal tissue, which has not been observed in response to most other proteins

Ruptura de la barrera hematoencefálica y síntesis intratecal de inmunoglobulinas II: síntesis local de inmunoglobulinas / Disruption of the blood- brain barrier and intrathecal synthesis of immunoglobulins II: local synthesis of immunoglobulins

Se aplican las fórmulas y gráficas de Reiber y de Reiber/Felgenhauer a un grupo de 75 pacientes pediátricos con infecciones del sistema nervioso central y convulsiones con fiebre, con el objetivo de conocer las características de su respuesta inmune intratecal en 2 sentidos: si hay síntesis local de inmunoglobulina A, M y G independientemente de las condiciones actuales del daño de la barrera hematoencefálica y su clasificación de acuerdo con la gráfica de Reiber. La mayor parte de los pacientes se ubicó en la zona 1 normal y en la zona 5 donde hay síntesis de IgG sin ruptura de la barrera; 2 pacientes con meningoencefalitis bacteriana y 1 viral se ubicaron en la zona 4 con síntesis de IgG y ruptura de la barrera y 1 paciente en la zona 2 con disfunción de la barrera. Según Reiber/Felgenhauer la mayoría de los pacientes sintetizaron IgA e IgG local la cual fue intensa en pacientes con meningoencefalitis bacterianas y meningococemia

Kinetics of specific IgM in patients of hepatic amoebiasis.

Entamoeba histolytica (EH) specific IgM was measured in 54 patients with diagnosed amoebic liver abscess (ALA), 13 with non-suppurative hepatic amoebiasis (NSHA) and 50 controls. The mean levels of EH specific IgM, estimated by ELISA were significantly raised in patients of invasive amoebiasis (both ALA and NSHA) compared to controls (P less than 0.05). EH specific IgG was also raised in both groups of patients. Follow up of patients with ALA showed a significant decline (P less than 0.05) in the specific IgM levels three months after treatment while the specific IgG antibodies persisted in high titres (1:160). Only four patients of NSHA could be followed up and all showed a decline in specific IgM levels. Raised specific serum IgM seems to be an indicator of active (invasive) amoebiasis.

Aplicación de una nueva fórmula en la síntesis intratecal de inmunoglobulinas / Implementations of a new formula in the intrathecal synthesis of immunoglobulins

Se aplicó la fórmula propuesta por Reiber y Felgenhauer a 462 pacientes pediátricos con meningoencefalitis. Se cuantificaron la IgA, IgM, IgG y la albúmina en suero y el líquido cefalorraquídeo por inmunodifusión radial. Hubo síntesis de IgA en el 58,87%, de IgM en el 57,79% y de IgC en el 23,8% de los pacientes estudiados. La cantidad de inmunoglobulinas sintetizadas con respecto al contenido total en el líquido cefalorraquídeo tuvo una alta variabilidad. En el caso de la IgM media local sintetizada fue mayor que en el resto de las inmunoglobulinas

Effect of suppressor cells on antibody producing cells in mice infected with Mycobacterium leprae.

Swiss albino mice were transfused with suppressor cells obtained after in vivo stimulation of mice with Con A (NS group). Some of the animals were infected with Mycobacterium leprae (NSI-group). Half of these animals were treated with dapsone (NSIT group). Adequate normal (NC) and infected (NI) controls were included. A plaque assay was carried out at different time periods to elucidate the effect of suppressor cells on antibody producing cells. No significant difference was seen in the number of plaque forming cells (PFC) in infected and dapsone treated animals (NSIT) when these were compared with controls. However significant increase seen in the number of IgM plaque forming cells at 6 months in NI and NSI groups and IgG PFC in NI group could be due to the peak footpad infection during this period. The significant decrease in the number of IgG PFC in NS and NSIT group compared to NC at 0 month is probably due to the suppressor cell activity in these groups.

Changes of Tetanus Specific IgG, IgM and IgG Subclasses after DPT Vaccination

We evaluated tetanus specific IgG, IgM, IgG subclasses after DPT vaccination in infants and children. Tetanus toxoid specific IgG, IgM IgG subclasses were measured to characterize the isotope profile of antibody against tetanus toxoid. The values of the tetanus specific IgG in the positive group were significantly increased compared to those of the control group, and were significantly increased after two inoculation. Tetanus specific IgG was very low in adults and neonates. In our tetanus specific IgG subclasses study, forty-five of 56 cases (80%) showed predominantly IgG1 antibody responses to tetanus toxoid, while twenty-five of 56 cases (45%) showed IgG4 responses. Both IgG1 and IgG4 responses were demonstrated in 17 cases (30%). So we suggest that IgG was mainly involved in humoral immune response after DPT vaccination, and IgG1 may play an important role among IgG subclasses. IgG4, alone or together with IgG1, can also play a role in immune response to tetanus toxoid.

Regulation of immunoglobulin secretion by T lymphocytes in human malaria.

In vitro studies were carried out on the nature of immunoglobulin synthesis and secretion by peripheral blood mononuclear leukocytes (PBMLs) and on the function of T and B cells from malaria patients. The mean values of secreted IgG and IgM concentrations of 22 malaria patient PMBLs were significantly higher than those of 20 normal PBMLs. When the suppressor T cell activity and the function of B cells in response to suppressor T cells were assayed by the cell co-culture technique, it was found that there was a decrease in suppressor T cell activity and the B cell function in response to normal suppressor T cells in malaria patients. The defects of these T and B cell functions may play some role in the immunological abnormalities seen in some malaria patients.

Assessment of antibody responses and protective immunity in cholera vaccinated subjects.

Cross-reactive antibody responses were assessed in volunteers vaccinated with classical Inaba and Ogawa cholera vaccines. The El Tor, Ogawa vibrios, the most often biotype, and serotype found to be the causative agent of cholera in Thailand, or their product were used throughout the in vitro and in vivo tests. The test involved were the passive hemagglutination test, vibriocidal tests and the mouse protection test. Classes of specific immunoglobulins produced in the volunteers were determined using anti-immunoglobulin enhancement of hemagglutination. It was found that the levels of hemagglutinating and vibriocidal antibodies reached their peaks on day 7 after the vaccination and were statistically constant for 3 months. Significant decrease was observed thereafter. The mouse protective antibody titer was highest at 1 month after the vaccination then declined significantly at the 6th month. Classes of specific immunoglobulins were found to be either IgM or IgG alone or mixture of both.