Obtenção de anticorpos para moleculas de expressão especifica em celulas natural Killer uterinas (NKu) de camundongos

AUTOR(ES)
DATA DE PUBLICAÇÃO

2001

RESUMO

During the pregnancy in the rodents a transient accumulation of natural killer (NK) cells 1s seen In the uterus. These cells found in the uterine environment express the surface antigen common to drculating NK (cNK) cells, but they do not show some cytokine receptors and do not trigger the cytoJytíc activity during normal pregnancy. Such a peculiar behavior of uNK ceH has not beeo fuUy explained aod their role io the immunoiogy of reproduction 1s stH! unc!ear. It has been reported the use of OBA (Dofichos bifJorus) lectin cytochernistry for selective staining of mouse uNK cel!. Since thís lectin does not react with othar !ymphocytes, the g!ycoconjugates reactive to OBA iectin localized on uNK cel! surface could be specific mo!ecules of such a cal! lineage. However, the glycoconjugates reactive to OBA lectin have no! been isolated ar characterized yet. The present work was proposed to obtain antibodies specific agaínst mouse uNK cel! and iooking identify mo!ecules of specific expression for. In this aim, the whole uNK ce!ls isolated from pregnant mice uterus were used as antigen and male mice of sarne strain, as host. From the animal producing antibodies against mouse uNK cell were collected the spleen and further processed to isolate the Iymphoeytes dones producing the monoclonal antibody (mAb) and the hybridomas of four clones were producad the ascites. The specificities of antibodies were evaluated by immunocytochemistry and immunoblotting. Only one among six immunized miee produced antibody against mouse uNK eells evaluated by immunocytochemistry. Four dones showing two different JabeJing pattern by immunocytochemistry were chosen to obtain ascites. The two (2C6.2B9, 2C6.2E4) clones producing mAS against uNK ceU cytop!asm eontents showed one positive band (41,4 KDa) in the westem blot matched to the DBA lectin positive bands. The other two ciones (2C4.1C10 e 2C4.5A 10) producJng mAb reacting on ceil surface and cytoplasm contents showed two positive bands (52,0 KDa and 53,8 KDa) corresponding to OBA lectin positive bands. Therefore, the antibody production against uNK cells afier intra-specie inoculation is the first direct evidence showíng uNK cells as being a cel! !ineage specific of pregnancy, which express specific molecules recognízed as non-self by male of the sarne strains. Among the mAb obtained, those reactive to antigen localized on call surface could be used as specific ceH marker of uNK once recognize the same glycoconjugates reactive to DBA lectin tha! seems to be expressed only by this cell

ASSUNTO(S)

celulas killer imunoglobulinas imunoglobulinas - reprodução

ACESSO AO ARTIGO

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