Jerold G. Woodward

Cloning and identification of the H-2Dp gene

We have cloned six different class I genes from a B10.P sperm library. After cotransfection with the herpes simplex tk gene, one L-cell line was found to react with six H-2Dp-specific monoclonal antibodies. The cell line L12a did not react with Kp-specific monoclonal antibodies. This identification was confirmed by mapping a 2.5 kb Bam H 1 restriction fragment present in the λ12a DNA clone to the D-TL region of H-2p. Only a single 8.8 kb Barn H1 fragment can be assigned to Kp by restriction fragment length polymorphism, while many others map to the D-TL interval. A restriction map of λ12a is presented.

Functional characterization of the H-2Dp gene product

A λ clone containing the Dp gene was used to transform L cells. The Dp product expressed was identified by two-dimensional gel electrophoreis and flow cytometry. The Dp product expressed by the L cells was recognized by DP-specific flow cytometry. The Dp product expressed by the L cells was recognized by Dp-specific but not Kp-specific killer T cells. This killing was inhibited by monoclonal antibodies specific for Dp but not Kp or Kk antigens. Similarly, lymphocytic choriomeningitis virus (LCMV) killer T cells from B 10.P mice were able to kill LCMV-infected L12a cells, but not LCMV-infected Ltk+. Again only Dp monoclonal antibodies could inhibit this killing.

Low-density mononuclear cells: Potent stimulators of the human MLR

We have applied purification strategies similar to those used to purify murine dendritic cells to human peripheral blood in an attempt to enrich for stimulators of the human mixed lymphocyte reaction (MLR). Equilibrium density centrifugation of peripheral blood mononuclear cells yields a population of low density cells that are potent stimulators of a human MLR. The stimulator cells are Dr+, nonlymphocytic, and weakly adherent. Strongly adherent monocytes, also present in the low-density cell population, do not stimulate a human MLR. This contrasts with other human MLR studies that ascribe stimulatory activity to adherent monocytes, and it indicates functional and morphological heterogeneity among monocytes.

Fine Specificity and Genetic Restriction of T Cell Clones Specific for Mouse Hepatitis Virus, Strain JHM

Mouse hepatitis viruses are members of the coronavirus group of animal viruses. Although named for their propensity to induce acute hepatitis in animals stressed by a variety of conditions, it has become clear that as a group they possess the ability to cause a diverse group of diseases in their natural host (Wege, et al, 1982). One strain of MHV, named JHM virus (JHMV), was isolated from mice found to have demyelinated lesions of the central nervous system (Bailey, et al, 1949). More recently it has been found that JHMV is not only capable of causing acute encephalomyelitis with demyelination but also chronic demyelination probably due to the establishment of a latent infection of oligodendroglia, the cells of myelin within the central nervous system (Herndon, et al, 1975; Stohlman and Weiner, 1981).

BIOLOGICAL PROPERTIES OF CLASS I MHC MOLECULES EXPRESSED AFTER DNA-MEDIATED GENE TRANSFER

ABSTRACT Mouse L cells ( H-2 k ) transfected with a genomic clone containing the gene for transplantation antigen H-2L express the H-2L d molecule on the cell surface in a form indistinguishable from the native molecules. To determine the applicability of this system to the study of major histocompatibility complex (MHC) gene function, several biological properties of a cloned H-2L d transformant, 8-5, were evaluated. When 8-5 cells were used as an immunogen in C3H ( H-2 k ) mice, an antiserum was produced with activity predominantly directed against H-2L d specificities. This illustrates the utility of this approach for defining previously uncharacterized class I gene products expressed in transformed cells. The H-2L d molecules expressed on 8-5 cells also function as direct target antigens and restricting elements for H-2L d specific and lymphocytic choriomeningitis virus specific cytotoxic T lymphocytes respectively. In both instances, recognition of 8-5 cells by effector T cells was blocked by addition of H-2L d specific antisera or monoclonal antibodies.