New insights of the local immune response against both fertile and infertile hydatid cysts.

BACKGROUND: Cystic echinococcosis is caused by the metacestode of the zoonotic flatworm Echinococcus granulosus. Within the viscera of the intermediate host, the metacestode grows as a unilocular cyst known as hydatid cyst. This cyst is comprised of two layers of parasite origin: germinal and laminated layers, and one of host origin: the adventitial layer, that encapsulates the parasite. This adventitial layer is composed of collagen fibers, epithelioid cells, eosinophils and lymphocytes. To establish itself inside the host, the germinal layer produces the laminated layer, and to continue its life cycle, generates protoscoleces. Some cysts are unable to produce protoscoleces, and are defined as infertile cysts. The molecular mechanisms involved in cyst fertility are not clear, however, the host immune response could play a crucial role. METHODOLOGY/PRINCIPAL FINDINGS: We collected hydatid cysts from both liver and lungs of slaughtered cattle, and histological sections of fertile, infertile and small hydatid cysts were stained with haematoxylin-eosin. A common feature observed in infertile cysts was the disorganization of the laminated layer by the infiltration of host immune cells. These infiltrating cells eventually destroy parts of laminated layer. Immunohistochemical analysis of both parasite and host antigens, identify these cells as cattle macrophages and are present inside the cysts associated to germinal layer. CONCLUSIONS/SIGNIFICANCE: This is the first report that indicates to cell from immune system present in adventitial layer of infertile bovine hydatid cysts could disrupt the laminated layer, infiltrating and probably causing the infertility of cyst.

Bovine (Bos taurus) humoral immune response against Echinococcus granulosus and hydatid cyst infertility.

Echinococcus granulosus, the agent of hydatid disease, presents an indirect life cycle, with canines (mainly dogs) as definitive hosts, and herbivores and human as intermediary ones. In intermediary hosts fertile and infertile cysts develop, but only the first ones develop protoscoleces, the parasite form infective to definitive hosts. We report the presence of bovine IgGs in the germinal layer from infertile cysts (GLIC), in an order of magnitude greater than in the germinal layer from fertile cysts (GLFC). When extracted with salt solutions, bovine IgGs from GLIC are associated with low or with high affinity (most likely corresponding to non specific and antigen specific antibodies, respectively). Specific IgGs penetrate both the cells of the germinal layer and HeLa cultured cells and recognize parasitic proteins. These results, taken together with previous ones from our laboratory, showing induction of apoptosis in the germinal layer of infertile hydatid cysts, provide the first coherent explanation of the infertility process. They also offer the possibility of identifying the parasite antigens recognized, as possible targets for immune modulation.

Antiophidian sera sterility control: topics in perspective / Controle de esterilidade de soros antiofídicos: temas em perspectiva

The aim of this work is to review the most important topics about the antiophidic sera sterility, including obtaining methods, sterilization procedures and clean room control using Vital Brazil Institute (VBI) as an example. Bibliographical research was performed through Medline, Lilacs, PubMed, ISI and the Fundação Oswaldo Cruz - RJ and VBI Libraries, from 1960 to 2009. The antiophidic sera for human use are immunobiologic products produced in Brazil by three national laboratories, including VBI. Due to the parenteral use, these products should be sterile and pyrogen-free, which demands the microbiological control during the whole fabrication process. The sterility and pyrogen tests are important steps to ensure the quality and safety of these immunobiological products. Thus, these tests are target for continue evaluation and improvement. The most interfering aspects in the consistency and analytical patterns include the proper method selection, sampling, culture conditions and validation criteria. As the national and international legal requirements are cautious with the assays validation and approval of sterile parenteral products; the intrinsic limitations for established assays still require more investigation aiming the continue improvement of the microorganism and contaminants detection methods and optimization of the analysis extent.

O objetivo deste trabalho é revisar os tópicos mais relevantes para o controle da esterilidade de soros antiofídicos, abordando-se métodos de obtenção, procedimentos de esterilização e o controle de áreas limpas utilizando como exemplo os procedimentos adotados pelo Instituto Vital Brazil (IVB). Um levantamento bibliográfico foi realizado no Medline, ISI, Biblioteca da Fundação Oswaldo Cruz-RJ e IVB, no período de 1960 a 2009. Os soros antiofídicos para uso humano são produtos imunobiológicos fabricados no Brasil por três laboratórios nacionais, dentre eles o IVB. Por serem de administração parenteral, devem ser obrigatoriamente estéreis e apirogênicos, exigindo controle microbiológico durante todo o processo de fabricação. O teste de esterilidade e apirogenia são importantes instrumentos para garantir a qualidade e segurança microbiológica desses produtos, sendo alvo de avaliações constantes para seus aprimoramentos. Os aspectos que mais interferem na sua consistência e valor analítico incluem a escolha adequada do método, amostragem, condições de cultivo e critérios de validação. À medida que os requisitos legais nacionais e internacionais se mostram rigorosos na validação de ensaios e aprovação de produtos estéreis parenterais, limitações intrínsecas ao ensaio padronizado requisitam mais investigações, objetivando o aprimoramento contínuo nos métodos de detecção de microorganisms, contaminantes e otimização do tempo total de análise.

Antiacrosin antibodies and infertility. II. Gene immunization with human proacrosin to assess the effect of immunity toward proacrosin/acrosin upon protein activities and animal fertility.

OBJECTIVE: To assess the effect of antiacrosin antibodies upon proacrosin/acrosin activities and animal fertility. DESIGN: Prospective study. SETTING: Basic research laboratory. PATIENT(S): A gene immunization (GI) model was developed; mice were injected with the sequence encoding human proacrosin (h-proacrosin), cloned in an expression vector. INTERVENTION(S): Subcloning of h-proacrosin in a eukaryotic expression vector (promoter, CMV; leader sequence, alpha-1 antitrypsin; pSF2-Acro); GI of female mice with this plasmid. MAIN OUTCOME MEASURE(S): The following parameters were evaluated: [1] adequate conditions for GI protocols, [2] humoral response to GI with pSF2-Acro, [3] protein regions recognized by the antibodies, and [4] effect of antibodies upon proacrosin/acrosin-ZPA binding and amidase activity, and animal fertility. RESULT(S): Conditions of female mice GI with the proacrosin sequence were established (plasmid purification with anion exchange chromatography and 40 microg of pSF2-Acro per dose) to trigger an immune response, reaching maximum levels at week 9 after the first injection. Antibodies produced by GI recognized human and mouse sperm acrosin systems, inhibited human proacrosin/acrosin interaction with recombinant human ZPA and protease activity, and negatively affected mouse IVF and early embryonic development. In addition, mice immunized with SF2-Acro exhibited a significantly lower size of fetuses. CONCLUSION(S): Antiacrosin antibodies developed by using GI inhibit human proacrosin/acrosin activities and impair mouse fertility.

Imunologia reprodutiva na prática clínica: uma visão crítica / Reproductive immunology in clinical practice: a critical view

Atualmente, grande importância tem sido dada aos aspectos imunológicos da reprodução humana. Embora a maior parte dos mecanismos imunológicos envolvidos na gestação não seja inteiramente compreendida, muitos casais inférteis têm sido submetidos à investigação imunológica e diferentes tipos dos assim chamados tratamentos imunológicos têm sido amplamente prescritos. Os exames que devem ser incluídos nesta investigação, bem como a eficácia da maioria dos tratamentos ainda não estão bem estabelecidos. O objetivo deste artigo é discutir o que se sabe sobre o real papel do sistema imune em casos de infertilidade e apresentar uma visão crítica da imunologia reprodutiva na prática clínica.

Anticuerpos antiespermatozoides en parejas con infertilidad de causa no explicada / Antisperm antibodies in couples with infertility of unknown origin

Se estudió una muestra de 75 parejas infértiles en relación con la presencia de anticuerpos antiespermatozoides en plasma seminal, moco cervical o adheridos a los espermatozoides, mediante la prueba de inmunobeads y se determinó la capacidad inhibidora del plasma seminal sobre la proliferación linfocitaria, para evaluar la capacidad inmunomoduladora del semen como participante en el mecanismo patogénico primario de aparición de los anticuerpos antiespermatozoides. Se organizaron las parejas en 2 grupos, según la presencia o no de anticuerpos antiespermatozoides y se compararon en cuanto a la frecuencia de embarazo y la capacidad inmunosupresora del plasma seminal. Además se comparó la incidencia de embarazo en grupos de parejas con capacidad inmunosupresora y sin ella. Se observó que la presencia de anticuerpos antiespermatozoides fue menos frecuente que la reportada por otros autores y no estuvo relacionada con la capacidad inmunosupresora del plasma seminal, tampoco influyeron sobre la frecuencia de embarazo la presencia de anticuerpos antiespermatozoides y la capacidad inmunosupresora del plasma seminal (AU)

Anticuerpos antiespermatozoides en parejas con infertilidad de causa no explicada / Antisperm antibodies in couples with infertility of unknown origin

Se estudió una muestra de 75 parejas infértiles en relación con la presencia de anticuerpos antiespermatozoides en plasma seminal, moco cervical o adheridos a los espermatozoides, mediante la prueba de inmunobeads y se determinó la capacidad inhibidora del plasma seminal sobre la proliferación linfocitaria, para evaluar la capacidad inmunomoduladora del semen como participante en el mecanismo patogénico primario de aparición de los anticuerpos antiespermatozoides. Se organizaron las parejas en 2 grupos, según la presencia o no de anticuerpos antiespermatozoides y se compararon en cuanto a la frecuencia de embarazo y la capacidad inmunosupresora del plasma seminal. Además se comparó la incidencia de embarazo en grupos de parejas con capacidad inmunosupresora y sin ella. Se observó que la presencia de anticuerpos antiespermatozoides fue menos frecuente que la reportada por otros autores y no estuvo relacionada con la capacidad inmunosupresora del plasma seminal, tampoco influyeron sobre la frecuencia de embarazo la presencia de anticuerpos antiespermatozoides y la capacidad inmunosupresora del plasma seminal(AU)

The 60 kDa heat shock protein in human semen: relationship with antibodies to spermatozoa and Chlamydia trachomatis.

The presence of the 60 kDa heat shock protein (hsp60) in seminal fluid and its relationship to sperm autoimmunity or a localized immune response to Chlamydia trachomatis were examined. Semen from 64 male partners of infertile couples with no history of a chlamydial infection were investigated. Hsp60 was identified by an enzyme-linked immunosorbent assay (ELISA) using a monoclonal anti-hsp60 antibody bound to wells of a microtitre plate and a polyclonal anti-hsp60 antibody for detection. Antisperm antibodies on motile spermatozoa were detected by immunobead binding, while antichlamydial immuno-globulin (Ig) A and IgG in seminal fluid were identified by a commercial ELISA (SeroELISA: Savyon Diagnostics, Beer-Sheva, Israel). RNA was purified from isolated seminal round mononuclear cells and tested for hsp60-specific mRNA by a reverse transcription polymerase chain reaction ELISA. Hsp60 was present in semen from nine (14.1%) men, 12 (18.8%) men had antisperm autoantibodies. 16 (25.0%) were positive for antichlamydial IgA and 17 (26.6%) had detectable hsp60-specific mRNA. The presence of hsp60 in semen correlated with the occurrence of antichlamydial IgA (P = 0.0005), hsp60 mRNA (P = 0.04) and antisperm antibodies (P = 0.05). Thus, hsp60 was present in a soluble form in semen primarily in men with evidence of immune system activation within their genital tract. The role of hsp60 in promoting or inhibiting immune responses within the genital tract remains to be determined.

Aspectos imunológicos da infertilidade conjugal / Immunological aspects of human infertility

Na infertilidade conjugal, apesar do avanço tecnológico, nao houve grande melhora nas taxas de gestaçao à termo. Uma lacuna existe no conhecimento sobre os mecanismos inerentes ao processo reprodutivo. Assim, atençao especial tem sido dada ao papel do sistema imunológico na reproduçao, uma vez que o feto deverá suscitar uma resposta imune no organismo materno. Os fatores que seriam responsáveis pela manutençao da gestaçao continuam sendo investigados. Apesar da fisiologia deste processo nao ser bem conhecida, alguns métodos diagnósticos têm sido empregados com o intuito de melhorar o prognóstico destes casais. Estes métodos trouxeram algumas contribuiçoes, mas há de se dizer que o processo reprodutivo é um sistema biológico complexo, que envolve o equilíbrio de vários sistemas orgânicos. Esta revisao procura mostrar aspectos atuais sobre este tema.

Autoimmunity to spermatozoa, asymptomatic Chlamydia trachomatis genital tract infection and gamma delta T lymphocytes in seminal fluid from the male partners of couples with unexplained infertility.

The relationship between an undetected, asymptomatic Chlamydia trachomatis genital tract infection, the concentration of gamma delta and alpha b T cells in semen and sperm autoimmunity was examined in 48 male partners of couples with unexplained infertility. Immunoglobulin A (IgA) antibodies to C. trachomatis were detected in seminal fluids from 14 (29.2%) of the men. Only four of these were positive for circulating anti-chlamydial IgA, suggesting that the stimulus for antibody production was within the genital tract. In contrast, four men were positive for anti-chlamydial IgG in their semen; all were also seropositive for anti-chlamydial IgG. T lymphocytes bearing the alpha beta and gamma delta antigen receptors were present in every semen sample. Men with seminal anti-chlamydial IgA, however, had significantly (P = 0.035) elevated semen gamma delta T cell concentrations (median 3100 cells/ml) than did men lacking this antibody (median 1400 cells/ml); concentrations of alpha beta T cells were comparable in both groups. Genital tract sperm autoimmunity, as shown by antibodies bound to motile ejaculated spermatozoa, was detected in 13 (27.1%) men. The presence of these antibodies was associated with elevated concentrations of both gamma delta (median 4200 versus 700 cells/ml) and alpha beta (median 5000 versus 850 cells/ml) T cells (P = 0.0002 and 0.0001 respectively). Men with antisperm antibodies only in their serum had seminal T cell concentrations comparable with men testing negative for antisperm antibodies. Anti-chlamydial IgA was identified in semen from four of 10 men with IgA bound to their spermatozoa and in none of the men with only spermatozoa-bound IgG.(ABSTRACT TRUNCATED AT 250 WORDS)

Factores embriotóxicos tempranos como posible mecanismo de infertilidad en pacientes con aborto habitual y esterilidad sin causa aparente / Embryologic toxic factors as infertility cause in patients habitual abortions with sterility without apparent cause

El objetivo del presente estudio es identificar la presencia de factores embriotóxicos tempranos como causa de infertilidad. Se estudia 15 sueros de pacientes divididos en dos grupos: de abortos habituales y con esterilidad sin causa aparente. Se realizó cultivo de embriones preimplantacionales de ratón que permite identificar en el suero de mujeres agentes embriotóxicos y mutagénicos incompatibles con el desarrollo embrionario.

Infertilidad inmunológica / Immunological infertility

Hasta el 30 por ciento de las infertilidades inexplicadas, tienen origen inmunológico. La infertilidad inmunológica agrupa una serie de trastornos en los que solamente ahora, se puede contar con la tecnología adecuada para diagnosticarla. Esta revisión comprende la fisiopatología de la formación de los anticuerpos anti-esperma, tanto en el hombre como en la mujer, y las técnicas para hacer un mejor enfoque diagnóstico de esta compleja enfermedad, sobre todo en nuestro medio, haciendo énfasis en la prueba de penetración del moco cervical in vitro. También se revisan los tratamientos que en la actualidad ofrece la medicina...

Anticuerpos ABH en moco cervical / Antybodies ABH in cervical mocus

Se estudian anticuerpos anti-A y anti-B en moco cervical pre y postratamiento con solución de 2-mercaptoetanol, utilizado para romper puentes disulfuro intracatenario de los anticuerpos IgM

Anticuerpos ABH en moco cervical / Antybodies ABH in cervical mocus

Se estudian anticuerpos anti-A y anti-B en moco cervical pre y postratamiento con solución de 2-mercaptoetanol, utilizado para romper puentes disulfuro intracatenario de los anticuerpos IgM (AU)

Anticuerpos antiespermatozoide en la infertilidad / Anti-spermatozoa antibodies in infertility

Utilizando la tecnica de macro-aglutinacion en gelatina propuesta por Kibrick, se estudio la presencia de anticuerpos antiespermatozoide en parejas infertiles. De 52 parejas, 33 consultaron por infertilidad primaria (63.6%) y 19 por secundaria (36.5%). El 46.1% de todas estas mujeres presentaron anticuerpos circulantes a titulos de 1:16 a 1:128, contra solo 3.8% en el grupo control. La frecuencia de anticuerpos circulantes por infertilidad primaria que en aquellas que lo hicieron por infertilidad secundaria (36.8). Solo 12 hombres dentro del grupo estudiado, presentaron anticuerpos a titulos menores o iguales a 1:16; no hubo en ellos correlacion ninguna con la calidad de los espermatozoides. No se encontro ninguna relacion entre la patologia gineco-obstetrica y la presencia de los anticuerpos circulantes