Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1986 Apr;52(1):36–44. doi: 10.1128/iai.52.1.36-44.1986

Flow cytometric analysis of the effects exerted by monoclonal antibodies on binding and uptake of Leishmania mexicana subsp. mexicana promastigotes by murine peritoneal macrophages.

K M Williams, J B Sacci, R L Anthony
PMCID: PMC262194  PMID: 3514462

Abstract

A flow cytometric assay was developed to quantitate the binding kinetics of Leishmania mexicana subsp. mexicana promastigotes to murine peritoneal macrophages and to determine if selected membrane-specific monoclonal antibodies would exert an effect on the binding process. A total of three monoclonal antibodies, all reactive with a similar 42-kilodalton surface membrane component by Western blot analysis, enhanced parasite-macrophage binding at levels greater than 45%. An additional three monoclonal antibodies that identified low-molecular-weight antigens of the promastigote (15 to 20 kilodaltons) had no effect on the binding process. Of these antibodies, however, one did appear to inhibit the internalization of the parasites after it attached to the macrophage membrane.

Full text

PDF
36

Images in this article

38Image
on p.38
38Image
on p.38
40Image
on p.40
41Image
on p.41
42Image
on p.42

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anthony R. L., Williams K. M., Sacci J. B., Rubin D. C. Subcellular and taxonomic specificity of monoclonal antibodies to New World Leishmania. Am J Trop Med Hyg. 1985 Nov;34(6):1085–1094. doi: 10.4269/ajtmh.1985.34.1085. [DOI] [PubMed] [Google Scholar]
  2. Bassøe C. F., Laerum O. D., Glette J., Hopen G., Haneberg B., Solberg C. O. Simultaneous measurement of phagocytosis and phagosomal pH by flow cytometry: role of polymorphonuclear neutrophilic leukocyte granules in phagosome acidification. Cytometry. 1983 Nov;4(3):254–262. doi: 10.1002/cyto.990040311. [DOI] [PubMed] [Google Scholar]
  3. Berman J. D., Dwyer D. M., Wyler D. J. Multiplication of Leishmania in human macrophages in vitro. Infect Immun. 1979 Oct;26(1):375–379. doi: 10.1128/iai.26.1.375-379.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bjerknes R. Flow cytometric assay for combined measurement of phagocytosis and intracellular killing of Candida albicans. J Immunol Methods. 1984 Aug 3;72(1):229–241. doi: 10.1016/0022-1759(84)90451-4. [DOI] [PubMed] [Google Scholar]
  5. Bray R. S. Leishmania mexicana mexicana: attachment and uptake of promastigotes to and by macrophages in vitro. J Protozool. 1983 May;30(2):314–322. doi: 10.1111/j.1550-7408.1983.tb02922.x. [DOI] [PubMed] [Google Scholar]
  6. Bruck C., Portetelle D., Glineur C., Bollen A. One-step purification of mouse monoclonal antibodies from ascitic fluid by DEAE Affi-Gel blue chromatography. J Immunol Methods. 1982 Sep 30;53(3):313–319. doi: 10.1016/0022-1759(82)90178-8. [DOI] [PubMed] [Google Scholar]
  7. Chang K. P. Antibody-mediated inhibition of phagocytosis in Leishmania donovani-human phagocyte interactions in vitro. Am J Trop Med Hyg. 1981 Mar;30(2):334–339. doi: 10.4269/ajtmh.1981.30.334. [DOI] [PubMed] [Google Scholar]
  8. Chang K. P. Cellular and molecular mechanisms of intracellular symbiosis in leishmaniasis. Int Rev Cytol Suppl. 1983;14:267–305. [PubMed] [Google Scholar]
  9. Chang K. P., Fong D. Cell biology of host-parasite membrane interactions in leishmaniasis. Ciba Found Symp. 1983;99:113–137. doi: 10.1002/9780470720806.ch7. [DOI] [PubMed] [Google Scholar]
  10. Chang K. P. Leishmania donovani-macrophage binding mediated by surface glycoproteins/antigens: characterization in vitro by a radioisotopic assay. Mol Biochem Parasitol. 1981 Nov;4(1-2):67–76. doi: 10.1016/0166-6851(81)90030-x. [DOI] [PubMed] [Google Scholar]
  11. Chang K. P. Leishmania donovani: promastigote--macrophage surface interactions in vitro. Exp Parasitol. 1979 Oct;48(2):175–189. doi: 10.1016/0014-4894(79)90097-3. [DOI] [PubMed] [Google Scholar]
  12. Ezekowitz R. A., Bampton M., Gordon S. Macrophage activation selectively enhances expression of Fc receptors for IgG2a. J Exp Med. 1983 Feb 1;157(2):807–812. doi: 10.1084/jem.157.2.807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Farah F. S., Samra S. A., Nuwayri-Salti N. The role of the macrophage in cutaneous leishmaniasis. Immunology. 1975 Oct;29(4):755–764. [PMC free article] [PubMed] [Google Scholar]
  14. Fortier A. H., Hoover D. L., Nacy C. A. Intracellular replication of Leishmania tropica in mouse peritoneal macrophages: amastigote infection of resident cells and inflammatory exudate macrophages. Infect Immun. 1982 Dec;38(3):1304–1308. doi: 10.1128/iai.38.3.1304-1308.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Handman E., Goding J. W. The Leishmania receptor for macrophages is a lipid-containing glycoconjugate. EMBO J. 1985 Feb;4(2):329–336. doi: 10.1002/j.1460-2075.1985.tb03633.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Handman E., Greenblatt C. L., Goding J. W. An amphipathic sulphated glycoconjugate of Leishmania: characterization with monoclonal antibodies. EMBO J. 1984 Oct;3(10):2301–2306. doi: 10.1002/j.1460-2075.1984.tb02130.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Handman E., Hocking R. E. Stage-specific, strain-specific, and cross-reactive antigens of Leishmania species identified by monoclonal antibodies. Infect Immun. 1982 Jul;37(1):28–33. doi: 10.1128/iai.37.1.28-33.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Herman R. Cytophilic and opsonic antibodies in visceral leishmaniasis in mice. Infect Immun. 1980 May;28(2):585–593. doi: 10.1128/iai.28.2.585-593.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Köhler G., Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495–497. doi: 10.1038/256495a0. [DOI] [PubMed] [Google Scholar]
  20. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  21. Leslie R. G. Immunoglobulin and soluble immune complex binding to phagocyte Fc receptors. Biochem Soc Trans. 1984 Oct;12(5):743–746. doi: 10.1042/bst0120743. [DOI] [PubMed] [Google Scholar]
  22. Leslie R. G., Niemetz A. H. Species specificity in the binding of IgG to macrophages. Immunology. 1979 Aug;37(4):835–840. [PMC free article] [PubMed] [Google Scholar]
  23. Merino F., Ajjam E., Hernández A., Dawidowicz K., Merino E. J. In vitro infection of murine macrophage by Leishmania brasiliensis. Mechanism of penetration. Int Arch Allergy Appl Immunol. 1977;55(1-6):487–495. doi: 10.1159/000231962. [DOI] [PubMed] [Google Scholar]
  24. Mosser D. M., Edelson P. J. Activation of the alternative complement pathway by Leishmania promastigotes: parasite lysis and attachment to macrophages. J Immunol. 1984 Mar;132(3):1501–1505. [PubMed] [Google Scholar]
  25. Nacy C. A., Diggs C. L. Intracellular replication of Leishmania tropica in mouse peritoneal macrophages: comparison of amastigote replication in adherent and nonadherent macrophages. Infect Immun. 1981 Oct;34(1):310–313. doi: 10.1128/iai.34.1.310-313.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Parham P. Monoclonal antibodies against HLA products and their use in immunoaffinity purification. Methods Enzymol. 1983;92:110–138. doi: 10.1016/0076-6879(83)92012-8. [DOI] [PubMed] [Google Scholar]
  27. Pearson R. D., Steigbigel R. T. Mechanism of lethal effect of human serum upon Leishmania donovani. J Immunol. 1980 Nov;125(5):2195–2201. [PubMed] [Google Scholar]
  28. Pearson R. D., Sullivan J. A., Roberts D., Romito R., Mandell G. L. Interaction of Leishmania donovani promastigotes with human phagocytes. Infect Immun. 1983 Apr;40(1):411–416. doi: 10.1128/iai.40.1.411-416.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Segal D. M., Hurwitz E. Binding of affinity cross-linked oligomers of IgG to cells bearing Fc receptors. J Immunol. 1977 Apr;118(4):1338–1337. [PubMed] [Google Scholar]
  30. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wyler D. J. In vitro parasite-monocyte interactions in human leishmaniasis. Evidence for an active role of the parasite in attachment. J Clin Invest. 1982 Jul;70(1):82–88. doi: 10.1172/JCI110606. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Zehavi U., El-On J., Pearlman E., Abrahams J. C., Greenblatt C. L. Binding of Leishmania promastigotes to macrophages. Z Parasitenkd. 1983;69(4):405–414. doi: 10.1007/BF00927696. [DOI] [PubMed] [Google Scholar]
  33. Zenian A. Leishmania tropica: biochemical aspects of promastigotes' attachment to macrophages in vitro. Exp Parasitol. 1981 Apr;51(2):175–187. doi: 10.1016/0014-4894(81)90106-5. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES