Related Experiment Videos
Expression of human CD4 by two human-mouse interlineage hybrids.
G M Taylor1, J E Morten, H Morten
1Department of Medical Genetics, St Mary's Hospital, Manchester, UK.
Journal of Immunogenetics
|August 1, 1988
Summary
Researchers created hybrid cell lines to study human CD4 expression. These cell lines, expressing CD4 (a key HIV target), are valuable tools for developing and analyzing anti-CD4 monoclonal antibodies.
Area of Science:
- Immunology
- Cell Biology
- Virology
Background:
- The CD4 molecule is a critical co-receptor for HIV entry into host cells.
- Developing effective anti-CD4 therapies requires robust tools for antibody development and analysis.
Purpose of the Study:
- To generate and characterize novel hybrid cell lines for studying human CD4 expression.
- To create a resource for the preparation, screening, and analysis of anti-CD4 monoclonal antibodies.
- To facilitate research into CD4 epitopes recognized by HIV.
Main Methods:
- Fusion of human B-lymphoid cells with mouse T-lymphoma BW5147 to create hybrid cell lines.
- Isolation of CD4+ and CD4- variants using fluorescence-activated cell sorting (FACS) with the OKT4 monoclonal antibody.
- Cytogenetic, isoenzyme, and DNA analysis to confirm chromosome presence and copy number.
- Screening of monoclonal antibodies against human T-cell antigens.
Main Results:
- Two hybrid cell lines, TF42 and TF53.1, expressing human CD4 were successfully generated.
- CD4 expression was confirmed to be associated with human chromosome 12, with multiple copies in TF42.
- Only antibodies targeting CD4 and Leu 8 reacted with CD4+ variants, indicating specificity.
Conclusions:
- The developed hybrid cell lines provide a valuable platform for anti-CD4 antibody research.
- These cell lines can aid in the development and validation of therapeutic and diagnostic antibodies.
- The resource supports investigations into HIV-CD4 interactions and epitope mapping.