The anti-Tac monoclonal antibody is known to bind to the p55 chain ofthe human interleukin 2 receptor and to inhibit proliferation of T cells by blocking interleukin 2 binding. However, use of anti-Tac as an immunosuppressant drug would be impaired by the human immune response against this murine antibody. We have therefore constructed a "humanized" antibody by combining the complementaritydetermining regions (CDRs) of the anti-Tac antibody with human framework and constant regions. The human framework regions were chosen to maximize homology with the anti-Tac antibody sequence. In addition, a computer model of murine anti-Tac was used to identify several amino acids which, while outside the CDRs, are likely to interact with the CDRs or antigen. These mouse amino acids were also retained in the humanized antibody. The humanized anti-Tac antibody has an affinity for p55 of 3 x 109 M-1, about

All human and murine immunoglobulin heavy chain variable region (VH) genes contain the sequence ATGCAAAT approximately 70 nucleotides 5' from the site of transcription initiation. This octanucleotide, in reverse orientation, is also found in all light chain variable region (VL) genes, and in the immunoglobulin heavy chain transcriptional enhancer. Transfection studies have established that this octamer is involved in the lymphoid-specific transcription of immunoglobulin genes. Octamer-containing fragments have been reported to bind a factor present in nuclear extracts of human cell lines; however, identical binding activity was detected in both B lymphoid and non-lymphoid cells. Here we establish that nuclear extracts from distinct cell types differ in their ability to interact with octamer-containing fragments. We have also detected a DNA-protein interaction that may be involved in the cell-type specificity of immunoglobulin expression, and we have determined that a sequence upstream of the octamer participates in an interaction with a nuclear protein(s).

The endogenous neuraminidase activity of various mouse lymphoid subpopulations and tissue compartments was examined by a sensitive fluorometric assay. These analyses indicated that activated T lymphocytes possessed a significantly higher level of intracellular neuraminidase than activated B or resting T or B lymphocytes. Examination of the level of neuraminidase in bone marrow, thymus, lymph node, and unfractionated spleen indicated that these lymphoid tissues contained significantly less neuraminidase than was detected in stimulated T cells. Kinetic studies revealed that the majority of the increase in neuraminidase activity occurred between 24 and 48 h following stimulation. Analysis of activated T lymphocytes prepared from a panel of inbred mouse strains indicated that cells from mice of the H-2v haplotype, which possess the Neu-1a allele and are deficient in liver neuraminidase, exhibited a level of activity which was significantly lower than that detected in stimulated T cells from other mouse strains. These results indicate that the endogenous neuraminidase activity of T lymphocytes increases upon stimulation, and that the level of this enzyme activity in lymphoid cells is also controlled by the Neu-1 locus, which is located in the H-2 region of the major histocompatibility complex.

No abstract