Biomedical subjects

Richard Longnecker

Publications and source records attributed to Richard Longnecker.

39 records · Page 3Linked to original sources

PubMed2001

Signal transduction through the B cell antigen receptor is normal in ataxia-telangiectasia B lymphocytes.

The rare human genetic disorder ataxia-telangiectasia (A-T) has multiple consequences including a variable degree of immunodeficiency. Khanna and co-workers (Khanna, K. K., Yan, J., Watters, D., Hobson, K., Beamish, H., Spring, K., Shiloh, Y., Gatti, R. A., and Lavin, M. F. (1997) J. Biol. Chem. 272, 9489-9495) evaluated signaling in Epstein-Barr virus (EBV) immortalized A-T lymphoblastoid cell lines (LCLs), derived from the B cells of A-T patients. They showed that A-T lymphoblastoid cells lack signaling through the B cell antigen receptor and concluded that the fault in A-T encompasses intracellular signaling in B cells. However, it is established that EBV latent membrane protein 2A (LMP2A) blocks signaling in EBV-bearing cells by interaction with cellular tyrosine kinases. To test whether the reported fault in A-T B cells was not inherent in A-T but the result of influence of wild-type EBV, we derived A-T LCLs with wild-type or LMP2A-deleted EBV and studied signaling in these cells in response to cross-linking the B cell antigen receptor. We report that intracellular calcium mobilization and tyrosine phosphorylation in LMP2A-depleted LCLs derived from A-T patients is indistinguishable from that in LMP2A-depleted LCLs derived from normal controls. Further, signaling is blocked similarly in A-T and normal lymphoblastoid cells bearing wild-type EBV. In conclusion there is no evidence of any defect in B cell receptor signal transduction in A-T B cells.

Ataxia Telangiectasia↗

PubMed2000

Epstein-Barr virus latent membrane protein 2A has no growth-altering effects when expressed in differentiating epithelia.

Previous studies using transgenic mice with B-cell expression of LMP2A demonstrated that LMP2A drives B-cell development and survival signal in the absence of normal B-cell receptor (BCR) signal transduction. To determine if LMP2A may have similar effects in epithelial differentiation, six transgenic murine lines were constructed and analysed with LMP2A expression directed to the epidermis by a keratin 14 (K14) promoter cassette. LMP2A protein expression was verified by immunofluorescence and immunoprecipitation of skin samples using LMP2A-specific antibodies. To evaluate the effects of LMP2A expression on epidermal differentiation, immunofluorescence and histochemistry were performed on tongue and tail samples of transgenic mice and their wild-type littermate controls using differentially expressed keratins. The analysis indicated that LMP2A does not alter the normal epithelial differentiation program in the epithelia of K14-LMP2A transgenic mice.

Animals↗

PubMed1999

Epstein-Barr virus lacking latent membrane protein 2 immortalizes B cells with efficiency indistinguishable from that of wild-type virus.

Epstein-Barr virus (EBV) is a human herpesvirus that efficiently transforms and immortalizes human primary B lymphocytes. In this study, the role of latent membrane protein 2 (LMP2) in EBV growth transformation was investigated. LMP2 is a virally encoded membrane protein expressed in EBV-immortalized B cells previously shown to be nonessential for EBV transformation. However, a recent study reported that LMP2 may be an important determinant for efficient B cell transformation (Brielmeier et al., Journal of General Virology 77, 2807-2818, 1996). In this study a deletion mutation was introduced into the LMP2 gene using an E. coli mini-EBV construct containing sufficient EBV DNA to result in growth transformation of primary B cells. In an alternative approach, the introduction of the gene encoding the enhanced green fluorescent protein (EGFP) by homologous recombination into the LMP2 gene of EBV strain B95-8, generating the same LMP2 deletion mutation is reported. Careful quantification of B cell transformation using the EGFP+ LMP2- recombinant virus determined that in liquid culture medium or in culture medium containing soft agarose there was no difference in the ability of LMP2- virus to immortalize primary human B cells when compared to that of wild-type virus.

Animals↗