Effect of 65 kDa Buffalo Placental Protein on B-Cell Proliferation and Antibody Response

  • Deepthi, K. (Animal Biochemistry Division, National Dairy Research Institute) ;
  • Rajput, Y.S. (Animal Biochemistry Division, National Dairy Research Institute)
  • Received : 2001.09.06
  • Accepted : 2001.12.04
  • Published : 2002.03.01


Immunosuppressive potential of 65 kDa buffalo placental protein (bPP65) on B-cell proliferation in vitro and antibody response in vivo was evaluated. B-cell proliferation was estimated by measuring incorporation of tritiated thymidine in buffalo lymphocytes while primary antibody responses against phytohaemagglutinin (PHA) or keyhole limpet haemocyanin (KLH) were evaluated in mice. bPP65 suppressed proliferation of lipopolysaccharide (a B-cell specific mitogen)-stimulated buffalo lymphocytes in vitro indicating suppression of B-cells. This suppression was dose dependent over the protein concentration range $25-100 {\mu}g/ml$. Primary antibody responses in mice against PHA and KLH in presence of bPP65 were lower as compared to in its absence but these were not statistically significant. Amino acid composition data of bPP65 and BSA suggested that bPP65 is different from BSA.


  1. Barman, H. K. and Y. S. Rajput. 1994. Inhibition of mouse $\times$ mouse hybridoma growth by milk and colostrum. Lait. 74:473-478.
  2. Bohn, H., W. Kraus and W. Winkler. 1982. New soluble placental tissue proteins: their isolation, characterisation, localisation and quantification. Placenta 4:67-81.
  3. Boyum, A. 1968. Isolation of mononuclear cells and granulocytes from human blood. Scand. J. Clin. Lab. Invest. 21(Suppl. 97) 77.
  4. Germain, G., T. Mignot, P. Bischof, M. Mouren and L. Cedard. 1989. Plasma levels of SP1-like immunoreactive material (SP1) and progesterone throughout pregnancy and following RU486-induced early abortion in the cynomolgus monkey (Macaca fascicularis). Human Reprod. 4:975-980.
  5. Pockley, A. G. and A. E. Bolton. 1990. The effect of human placental protein 14 (PP14) on the production of interleukin-1 from mitogenically stimulated mononuclear cell culture. Immunology 69:277-281.
  6. Segerson, E. C., R. J. Moffatt, F. W. Bazer and R. M. Roberts. 1984. Suppression of phytohemagglutinin-stimulated lymphocyte blastogenesis by ovine uterine milk protein. Biol. Reprod. 30:1175-1186.
  7. Tease, I. A., A. T. Fazleabas and W. Y. Chan. 1989. Characterisation of baboon pregnancy specific $\beta$-1-glycoprotein. Biol. Reprod. 41:1113-1121.
  8. Bradford, M. M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72:248-254.
  9. Harlow, E and D. Lane. 1988. Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, New York.
  10. Laemmili, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680-685.
  11. Hansen, P. J. 1997. Interactions between the immune system and the bovine conceptus. Theriogenology 47:121-130.
  12. Pandian, A. M. C., R. D. Lambert and R. Roy. 1988. Immunosuppressive effects of rabbit blastocoelic fluid and embryo culture medium. J. Reprod. Immunol. 13:221-234.
  13. Pockley, A. G. and A. E. Bolton. 1989. Placental protein 14 (PP14) inhibits the synthesisof interleukin-2 receptors from phytohaemagglutinin-stimulated lymphocytes. Clinical Expertl. Immunol. 77:252-256.
  14. Segerson, E. C. and F. W. Bazer. 1989. High molecular weight basic and acidic immunosuppressive protein components in uterine secretions of pregnant cows. Biol. Reprod. 41:1014.
  15. Smith, A. D. and J. E. Wilson. 1986. A modified ELISA that selectively detects monoclonal antibodies recognizing native antigen. J. Immunol. Methods 94:31-35.
  16. Mehta, S. and Y. S. Rajput. 2001. Purification and characterisation of an immunosuppressive protein from buffalo placenta. Buffalo J. (In press).
  17. Watson, E. D. 1990. Suppression of lymphocyte activity by culture supernatent from horse embryos and endometrium. Biol. Reprod. 42:294-300.
  18. Mehta, S. and Y. S. Rajput. 1999. Immunosuppressive activity in buffalo placenta. Indian J. Exptl. Biol. 37:959-964.
  19. Newton, G. R., P. J. Hansen and B. G. Low. 1988. Characterisation of a high molecular weight glycoprotein secreted by the preimplantation bovine conceptus. Biol. Reprod. 39:553-560.
  20. Roth, T. L., K. L. White, D. L. Thompson Jr., B. E. Barry, J. S. Capehart, D. R. Colborn and M. H. Rabb. 1990. Suppression of lymphocyte proliferation by a >30,000 molecular weight factor in horse conceptus conditioned medium. Biol. Reprod. 43:298-304.
  21. Murray, M. K., E. C. Segerson, P. J. Hansen, F. W. Bazer and R. M. Roberts. 1987. Suppression of lymphocyte activation by a high molecular weight glycoprotein released from preimplantation ovine and porcine conceptus. Am. J. Reprod. Immunol. Microbiol. 14:38-44.
  22. Veselsy, L., J. Dostal, V. Holan, J. Soucek and B. Zelezna. 1996. Effect of boar seminal immunosuppressive fraction on B lymphocytes and of primary antibody response. Biol Reprod. 55:194-199.
  23. Hau, J. 1986. Animal proteins immunologically cross-reacting with human pregnancy associated $\alpha$2-glycoprotein (PZP, $\alpha$ 2 PAG, PA $\alpha$ 2G). In: Pregnancy proteins in animals.(Ed. J.Hau). Walter de Gruyter-Berlin, New York, pp. 445-451.
  24. Hugli, T. E. 1989. Techniques in protein chemistry. Academic press, Inc. Sandiego, California.
  25. Hansen, P. J., D. C. Stephenson, B. G. Low and G. R. Newton. 1989. Modification of immune function during pregnancy by products of the sheep uterus and conceptus. J. Reprod. Fertil. 37:55-61.
  26. Tatarinov, Y. S. and V. N. Masyukevich. 1970. Immunochemical identification of new $\beta$-1 globulin in the blood serum of pregnant women. ByII. Eksp. Biol. Med. USSR. 69:66-68.
  27. Bohn, H., R. Johannsen and W. Kraus.1980. New placental protein (PP15) with immunosuppressive properties. Arch. Gynaecol. 230:167-172.
  28. Deepthi, K. 2000. Studies on effect of 65 kDa buffalo placental protein on B and T lymphocyte response. MSc. Thesis, National Dairy Research Institute Deemed University, Karnal, India.
  29. Hansen, P. J., E. C. Segerson and F. W. Bazer. 1987. Characterisation of immunosuppressive substances in the basic protein fractions of the uterine secretions from pregnant ewes. Biol. Reprod. 36:393-403.
  30. Newton, G. R., J. L. Vallet, P. J. Hansen and F. W. Bazer. 1989. Inhibition of lymphocyte proliferation of ovine trophoblast protein-1 and a high molecular weight glycoprotein produced by the preimplantation sheep conceptus. Am. J. Reprod. Immunol. Microbiol. 19:99-107.