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Studies on human T lymphocyte colonies


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Woods, Gregory Mark 1984 , 'Studies on human T lymphocyte colonies', PhD thesis, University of Tasmania.

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This thesis examined the ability of human T lymphocytes to
form colonies in semisolid agar.
A two step procedure requiring a continued supply of PHA, a
source of SRBC during the agar culture and a high agar concentration
to restrict the freely mobile proliferating cells was developed for
the culturing of human lymphocytes in semisolid agar. The technique
was later modified by replacing PHA with an IL2-containing lymphokine
during the agar phase. The T cell nature of the colonies was confirmed
by their ability to form E-rosettes. An absence of colonies from
E-rosette depleted cells compared to E-rosette enriched cells
strengthened this finding and indicated that the colony forming cells
were T cells. The requirement for SRBC was investigated and it was
found that red cells from a variety of mammalian sources supported
colony formation through the elaboration of soluble factors which
diffused through the agar and did not require cell-cell contact for
their expression.
Experiments relating to the cellular interactions involved
in colony formation emphasised that cooperation between at least three
cell populations was essential. The interaction of these cells could
be described by two reactions. The first reaction required an adherent
cell (probably monocyte) population which, together with PHA, provided
the necessary signals for T cell activation. The activated T cells
interacted during the second or proliferative reaction. In response to
PHA, accessory T cells released IL2 which permitted colony forming
cells to proliferate. Thus PHA-induced T colony formation required the
cooperation of adherent cells for the activation reaction (T ->T') and IL2-producing T cells for the proliferative reaction (T' -> nT').
The microfilament/Microtubule network and the redistribution of
receptor :membrane complexes appeared to be an important feature of T
cell function with regard to both mitogen and IL2 binding.
Utilising the colony forming ability of T lymphocytes
studies on radiation induced mitotic death were carried out. Colony
formation was sensitive to low doses of irradiation although PHA
offered a limited degree of protection when cells were irradiated
prior to, or in the early stages of activation. This protection could
be explained by activities of repair enzymes which coincided with
early cellular activation events.
The human T lymphocyte colony assay was found to be a
valuable in vitro model for studying the steps involved in T
lymphocyte activation and proliferation.

Item Type: Thesis - PhD
Authors/Creators:Woods, Gregory Mark
Keywords: Lymphocytes, T cells
Copyright Holders: The Author
Copyright Information:

Copyright 1984 the Author - The University is continuing to endeavour to trace the copyright owner(s) and in the meantime this item has been reproduced here in good faith. We would be pleased to hear from the copyright owner(s).

Additional Information:

Thesis (Ph.D) - University of Tasmania, 1984. Bibliography: leaves 142-152

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