Adhesion inhibition of Mycoplasma iowae to chicken lymphoma DT40 cells by monoclonal antibodies reacting with a 65-kD polypeptide.
Tissue- and cell-specific attachment of mycoplasmas is a key aspect of the host-parasite relationship. In this study, monoclonal antibodies (MAbs) recognizing surface membrane polypeptides with molecular masses of 46 kD (p46) and 65 kD (p65), respectively, were examined in a microtiter cell attachment (agglutination) inhibition assay. MAbs MI3, MI6, and MI12 reacting with p65 polypeptide of Mycoplasma iowae inhibited attachment of the organisms to chicken lymphoma (DT 40) cells. One MAb (MI2) that reacted with p65 in immunoblots did not inhibit cell attachment, possibly because of the intrinsic native conformation of the epitope(s) in intact mycoplasmas as opposed to the linear state (sodium dodecyl sulfate denatured) in immunoblots. More pronounced M. iowae adherence inhibition was demonstrated by polyclonal turkey and mouse anti-M. iowae antisera compared with MAbs. Immunogold labelling followed by electron microscopy allowed us to localize the MAb-recognized epitopes on the membrane surface of M. iowae. On the basis of the cell attachment inhibition of M. iowae by specific MAbs (MI3, MI6, and MI12), we propose that the p65 polypeptide plays a role in cytadherence. The ability of polyclonal antisera to inhibit attachment of M. iowae more efficiently than the MAbs suggests that additional epitopes within p65 and/or other proteins are involved in cell attachment.
Fiorentin
L
L
Department of Pathobiology, College of Veterinary Medicine, Auburn University, AL 36849-5516, USA.
Panangala
V S
VS
Zhang
Y
Y
Toivio-Kinnucan
M
M
D000818:Animals; D000911:Antibodies, Monoclonal; D001422:Bacterial Adhesion / Q000502:physiology*; D002462:Cell Membrane / Q000382:microbiology / Q000473:pathology / Q000648:ultrastructure; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry; D005260:Female; D008223:Lymphoma / Q000382:microbiology / Q000662:veterinary; D008565:Membrane Proteins / Q000737:chemistry / Q000276:immunology / Q000502:physiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D009174:Mycoplasma / Q000502:physiology* / Q000648:ultrastructure; D009363:Neoplasm Proteins / Q000737:chemistry / Q000276:immunology / Q000502:physiology*; D011201:Poultry Diseases / Q000382:microbiology; D011487:Protein Conformation; D014407:Tumor Cells, Cultured; D014422:Turkeys
D000911:Antibodies, Monoclonal; D000939:Epitopes; D008565:Membrane Proteins; D009363:Neoplasm Proteins