What question did this study set out to answer?

To determine whether murine T cell-replacing factor produced by the B151K12 hybridoma possesses intrinsic B cell growth factor II proliferative activity.

June 1, 1985The Journal of Immunology

BCGFII activity on activated B cells of a purified murine T cell-replacing factor (TRF) from a T cell hybridoma (B151K12).

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Authors

NHNobuyuki HaradaNippon Shokubai (Japan)YKY. KikuchiHokkaido University ATA TominagaKyoto University

Discussion

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Implication

In vitro study reveals dual growth and differentiation activity of T cell-replacing factor in activated B cells, indicating a single molecule mediates both immune functions.

Key Points

To determine whether murine T cell-replacing factor produced by the B151K12 hybridoma possesses intrinsic B cell growth factor II proliferative activity.
Assayed cell-free supernatants from murine B151K12 hybridomas for lymphokine activity, immunoglobulin secretion, and [3H]thymidine incorporation in dextran sulfate-stimulated normal B cells and BCL1 leukemia cells.
Purified active proteins sequentially using ammonium sulfate precipitation, DEAE-cellulose, Blue-Sepharose, hydroxylapatite, gel permeation fast protein liquid chromatography (FPLC), and reverse-phase FPLC.
Tested functional identity via cellular absorption using BCL1 cells and targeted neutralization using the B cell-specific monoclonal antibody 9T1.
T cell-replacing factor and growth factor activities co-purified across all fractionation steps, co-eluting at 44% to 48% acetonitrile on reverse-phase FPLC with an apparent molecular weight of 50 to 60 kDa (18 kDa under reducing SDS-PAGE).
Cellular absorption with BCL1 tumor cells simultaneously removed both differentiation and proliferative activities from the purified preparation.
Monoclonal antibody 9T1 inhibited both the differentiation-dependent plaque-forming cell response and growth factor-mediated proliferation in BCL1 cells.

Cite This Study

Harada et al. (1985) studied this question.

synapsesocial.com/papers/6a9837ba161d47660415fca0 https://doi.org/10.4049/jimmunol.134.6.3944

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June 1, 1985The Journal of Immunology

BCGFII activity on activated B cells of a purified murine T cell-replacing factor (TRF) from a T cell hybridoma (B151K12).

View Full Paper

Ask AI

Bookmark

Authors

NHNobuyuki HaradaNippon Shokubai (Japan)YKY. KikuchiHokkaido University ATA TominagaKyoto University

Discussion

Member takes

Implication

In vitro study reveals dual growth and differentiation activity of T cell-replacing factor in activated B cells, indicating a single molecule mediates both immune functions.

Key Points

To determine whether murine T cell-replacing factor produced by the B151K12 hybridoma possesses intrinsic B cell growth factor II proliferative activity.
Assayed cell-free supernatants from murine B151K12 hybridomas for lymphokine activity, immunoglobulin secretion, and [3H]thymidine incorporation in dextran sulfate-stimulated normal B cells and BCL1 leukemia cells.
Purified active proteins sequentially using ammonium sulfate precipitation, DEAE-cellulose, Blue-Sepharose, hydroxylapatite, gel permeation fast protein liquid chromatography (FPLC), and reverse-phase FPLC.
Tested functional identity via cellular absorption using BCL1 cells and targeted neutralization using the B cell-specific monoclonal antibody 9T1.
T cell-replacing factor and growth factor activities co-purified across all fractionation steps, co-eluting at 44% to 48% acetonitrile on reverse-phase FPLC with an apparent molecular weight of 50 to 60 kDa (18 kDa under reducing SDS-PAGE).
Cellular absorption with BCL1 tumor cells simultaneously removed both differentiation and proliferative activities from the purified preparation.
Monoclonal antibody 9T1 inhibited both the differentiation-dependent plaque-forming cell response and growth factor-mediated proliferation in BCL1 cells.

Cite This Study

Harada et al. (1985) studied this question.

synapsesocial.com/papers/6a9837ba161d47660415fca0 https://doi.org/10.4049/jimmunol.134.6.3944

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