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Isolation and Activation of Murine Lymphocytes
Published on: October 30, 2016
A simplified microtechnique for measuring human lymphocyte proliferation after stimulation with mitogen and specific
1Department of Parasitology, Hebrew University, Hadassah Medical School, Jerusalem, Israel.
Journal of Immunological Methods
|September 13, 1988
Summary
A new microtechnique allows lymphocyte proliferation testing for cutaneous leishmaniasis using only a finger prick of blood. This method simplifies diagnosis and enables large-scale epidemiological studies of the parasitic disease.
Area of Science:
- Immunology
- Parasitology
- Cell Biology
Background:
- Cutaneous leishmaniasis involves a significant cell-mediated immune response.
- Conventional in vitro proliferation assays require large blood volumes, limiting their use.
- A simpler method is needed for assessing cellular immunity in leishmaniasis.
Purpose of the Study:
- To develop and validate a microtechnique for measuring human lymphocyte proliferation.
- To enable lymphocyte proliferation studies using minimal blood volumes obtained via finger prick.
- To assess the feasibility of this microtechnique for epidemiological studies of cutaneous leishmaniasis.
Main Methods:
- A microtechnique using 100 microliters of blood from a finger prick was developed.
- Mononuclear cells were directly cultured without Ficoll separation.
- Lymphocyte responses to Leishmania major, PPD, and mitogen were compared to conventional methods.
Main Results:
- The microtechnique yielded comparable results to conventional Ficoll separation methods.
- Individuals cured of Leishmania major infection showed a robust response to the specific antigen.
- No significant difference was observed between the microtechnique and Ficoll separation.
Conclusions:
- This microtechnique simplifies the assessment of cell-mediated immunity in cutaneous leishmaniasis.
- The method is suitable for epidemiological studies due to its minimal blood requirement.
- It facilitates the study of immune responses in leishmaniasis patients and cured individuals.
