Detection of Retroviral Superantigen and Products of the Envelope Gene from Endogenous Mouse Mammary Tumor Virus in B Cells from BALB/c Mice

Minor lymphocyte-stimulating antigens and other superantigens have been shown to be encoded by the 3ʹ long terminal repeat (LTR) open reading frame (ORF) of the endogenous and exogenous mammary tumor viruses. We have previously reported the presence of an antigen(s) related to mouse mammary tumor virus (MMTV) env products in splenic B cells of BALB/c mice. By Western blots an MMTV-related molecule of 68 kDa was detected in splenic preparations of B lymphocytes, but not in T cells. Antibodies against the MMTV envelope proteins gp52 and gp36, obtained by elution after binding to nitrocellulose in the presence of purified MMTV, reacted in Western blots with a 68-kDa protein present in B cells, indicating that this molecule is related to both MMTV envelope proteins. Using antibody against the MMTV 3ʹ LTR ORF coding sequence, 10-15% of splenic B cells reacted by immunoperoxidase staining with this reagent, while no such staining was observed in splenic T cell preparations. Furthermore, in preparations of splenic B cells, but not T cells, two bands of 68 and 33 kDa, respectively, were detected by Western blots using the anti-ORF. These results demonstrate that the superantigen protein is present in B cells of BALB/c mice in two distinct forms, i.e., as a 68-kDa molecule, possibly associated with products of the env gene, and as a 33-kDa form.