Immunodetection of human T-cell lymphotropic virus type I core protein in biological samples by using a monoclonal antibody immunoassay

Papsidero, Lawrence D.; Swartzwelder, F; Sheu, M; Montagna, Richard A.; Ehrlich, GD; Bhagavati, Satyakam; Dosik, Harvey; Sninsky, JJ; Poiesz, Bernard J.

doi:10.1128/jcm.28.5.949-955.1990

Cited by 16 publications

(7 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Some target cells were maintained in continuous culture after infection for up to 30 days. Sequential aliquots were removed and analyzed for HTLV-I DNA by PCR and HTLV-I protein expression, including reverse transcriptase (18), p19 Gag (16), and gp46 Env (1Sa). b A total of 5 x 105 PBMC at day 3 following activation with PHA (10 jig/ml) and delectinized IL-2 (20%, vol/vol).…”

Section: Methodsmentioning

confidence: 99%

Infection of peripheral blood mononuclear cells and cell lines by cell-free human T-cell lymphoma/leukemia virus type I

et al. 1992

View full text Add to dashboard Cite

Previous studies of in vitro infection by human T-cell lymphoma/leukemia virus type I (HTLV-I) have required cocultivation of target cells with HTLV-I cell lines or vesicular stomatitis virus pseudotypes containing HTLV-I envelope proteins. We report here the development of a cell-free infection assay for HTLV-I. Target cells were incubated with purified, DNase-treated HTLV-I virions for 4 h at 37°C. Target cell DNA was then analyzed for the presence of newly synthesized HTLV-I proviral DNA by the highly sensitive polymerase chain reaction. Using this assay system, we have been able to consistently detect in vitro infection of a variety of cellular targets by different HTLV-I isolates. Optimal infection required the presence of 10 ,ug of DEAEdextran per ml. The assay was dose dependent with respect to virus input. In general, the amount of proviral DNA detected correlated with the level of HTLV-I receptors present on the surface of the target cells, as measured by fluorochrome-labelled HTLV-I binding. Finally, the specificity of the assay was confirmed by demonstrating that the cell line, Llq, a somatic cell hybrid containing human chromosome 17q, to which the gene for the HTLV-I receptor has been mapped, was susceptible to infection by HTLV-I, while the parental mouse cell line from which it was derived, LMTK-, which lacks human chromosome 17q, was not.

show abstract

Section: Methodsmentioning

confidence: 99%

Infection of peripheral blood mononuclear cells and cell lines by cell-free human T-cell lymphoma/leukemia virus type I

et al. 1992

View full text Add to dashboard Cite

show abstract

“…Cultures were maintained in complete RPMI 1640 medium supplemented with 10% IL-2. Culture supernatants were monitored at approximately weekly intervals for the presence of HTLV antigens by antigencapture assays (HTLV-I p19, Cellular Products, Buffalo; NY, Papsidero et al, 1990;and HTLV-1/11 p24, Coulter;Lairmore et al, 1990). Resultant absorbance values of both tests were compared with known standard curves of viral core antigens performed in the same assay.…”

Section: Detection Of Htlv-i In Pbmc and Tissue Culttiresmentioning

confidence: 99%

Comparative biological responses of rabbits infected with human T‐lymphotropic virus type I isolates from patients with lymphoproliferative and neurodegenerative disease

Lairmore¹,

Roberts²,

Frank³

et al. 1992

Intl Journal of Cancer

View full text Add to dashboard Cite

An experimental rabbit model was used to determine host responses to infection by various human T-lymphotropic virus type-I (HTLV-I) strains. Seven groups of 4 to 5 rabbits each were inoculated with lethally-irradiated HTLV-I-infected cell lines derived from patients with adult T-cell leukemia/lymphoma or from patients with HTLV-I-associated myelopathy. Four separate control groups of 2 rabbits each were inoculated with similarly prepared HTLV-I-negative cells derived from rabbits or humans. Anti-viral antibody responses were assessed by immunoblot assay and hematologic parameters were measured using automated cell counters and cytologic staining. The virologic status of challenged rabbits was determined by co-culture and HTLV-I antigen capture assay, as well as by polymerase chain reaction (PCR) amplification of HTLV-I DNA from peripheral blood mononuclear cells (PBMC) or tissues. The HTLV-I inocula could be separated into groups based upon their infectivity to rabbits: highly infectious strains elicited intense serologic responses and were detected frequently in tissues by antigen and PCR assays, while other strains were moderately to poorly infectious, induced weak antibody responses and were infrequently detected by antigen and PCR assays. Overall, PBMC appeared to have the greatest quantity of HTLV-I containing cells, while bone marrow was a poor source of virus. No clinical or hematologic abnormalities were evident during the 24-week course of infection. Taken together, our results suggest there is heterogeneity in the biological response to HTLV-I infection which is, in part, dependent on the infecting strain of virus.

show abstract

“…Cell-free culture supernatants were obtained at log phase of growth (0.8 x 106 to 1.2 x 106 cells per ml) by sedimentation of the cells at 5,000 x g; the supernatants were then filtered through 0.45-,um-pore-size filters. Viral cultures were done as described previously (38).…”

Section: Methodsmentioning

confidence: 99%

“…Measurements of HTLV-I p19 (viral matrix protein) were performed by antigen capture immunoassay, as described previously (38), with commercial reagents (Retro-Tek HTLV-I Antigen ELISA kit; Cellular Products, Inc.). This immunoassay is HTLV-I specific and does not cross-react with HTLV-II (38). Immunoreactivity of MAb 6C2 with recombinant and native viral antigens with which microwells were coated.…”

Section: Triton Bfomctengagmentioning

confidence: 99%

See 1 more Smart Citation

Monoclonal antibodies and chemiluminescence immunoassay for detection of the surface protein of human T-cell lymphotropic virus

Papsidero¹,

Dittmer²,

Vaickus³

et al. 1992

J Clin Microbiol

View full text Add to dashboard Cite

Monoclonal antibodies (MAbs) raised against human T-cell lymphotropic virus type I (HTLV-I) recognized five distinct antigenic domains of viral env gene-encoded proteins. By using recombinant env proteins and synthetic peptides as mapping antigens, it was determined that the most immunogenic region represented a central portion of the retroviral surface protein (domain 2; amino acids 165 to 191). However, only a single MAb was able to react

show abstract

Immunodetection of human T-cell lymphotropic virus type I core protein in biological samples by using a monoclonal antibody immunoassay

Cited by 16 publications

References 39 publications

Infection of peripheral blood mononuclear cells and cell lines by cell-free human T-cell lymphoma/leukemia virus type I

Infection of peripheral blood mononuclear cells and cell lines by cell-free human T-cell lymphoma/leukemia virus type I

Comparative biological responses of rabbits infected with human T‐lymphotropic virus type I isolates from patients with lymphoproliferative and neurodegenerative disease

Monoclonal antibodies and chemiluminescence immunoassay for detection of the surface protein of human T-cell lymphotropic virus

Contact Info

Product

Resources

About