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Isolation and Activation of Murine Lymphocytes
Published on: October 30, 2016
Reference intervals for functional lymphocyte proliferation studies using 3H thymidine uptake in adults
Rohan Ameratunga1, Danny Lim2, Hilary Longhurst3
1Department of Clinical Immunology, Auckland Hospital, Park Rd, Grafton, 1010 Auckland, New Zealand; Department of Virology and Immunology, Auckland Hospital, Park Rd, Grafton, 1010 Auckland, New Zealand; Department of Molecular Medicine and Pathology, Faculty of Medical and Health Sciences, University of Auckland, New Zealand.
Insights
This study establishes reference intervals for 3H thymidine uptake assays, a key test for cellular immunity. These intervals aid in diagnosing immunodeficiencies and guiding treatment decisions for patients.
Area of Science:
- Immunology
- Clinical Diagnostics
Background:
- Functional cellular immunity tests are crucial in clinical settings.
- 3H thymidine uptake assays offer high sensitivity for assessing T cell responses.
- Assessing cellular immunity is vital for diagnosing immunodeficiencies and managing vaccine efficacy.
Purpose of the Study:
- To establish reference intervals for 3H thymidine uptake assays.
- To provide benchmarks for evaluating functional cellular immunity in healthy adults.
- To support diagnostic laboratories in interpreting cellular immunity test results.
Main Methods:
- Reference intervals were calculated for over 250 healthy adults.
- Tests included responses to lectins, anti-CD3 (OKT3), and vaccine antigens (Candida, tetanus, diphtheria).
- Volunteer blood donors' samples served as controls for patient diagnostic tests.
Main Results:
- Control samples showed uniform response to PHA.
- Responses to other lectins, OKT3, and antigens were heterogeneous.
- Variability in toxoid responses correlated with donors' immunization status.
Conclusions:
- 3H thymidine uptake assays remain valuable due to sensitivity, despite non-radioactive alternatives.
- Reference intervals aid in diagnosing conditions like Severe Combined Immunodeficiency (SCID).
- Results inform clinical decisions regarding live attenuated viral vaccines and hematopoietic stem cell transplantation (HSCT).
Aims:
This study aimed to establish the reference intervals for the 3H thymidine uptake assays of functional cellular immunity. Functional tests of cellular immunity play a critical role in several clinical scenarios. An excellent T cell response to vaccine antigens is arguably the best in vitro test of normal cellular immunity.
Methods:
In this article the reference intervals for over 250 healthy adults were calculated for lectins, anti-CD3 (OKT3) and antigens comprising Candida, tetanus and diphtheria toxoids. These samples were provided by volunteer blood donors as controls to be run in parallel with diagnostic tests of in vitro cellular immunity in patients with suspected immunodeficiency.
Results:
The control samples uniformly responded to PHA. The response to other lectins, OKT3 and antigens was more heterogeneous. The variable responses to diphtheria and tetanus toxoids likely reflected the immunization status of these blood donors to these vaccine antigens.
Conclusions:
Although there has been a recent move to implement non-radioactive methods for functional cellular immunity, 3H thymidine uptake continues to be utilized in many diagnostic laboratories because of excellent sensitivity. Robust vaccine-induced cellular responses in immunodeficient patients may inform clinical decisions such as using live attenuated viral vaccines. Similarly, markedly impaired responses to phytohemagglutinin A (PHA), in the appropriate clinical context, supports a diagnosis of Severe Combined Immunodeficiency (SCID) and the need for urgent hematopoietic stem cell transplantation (HSCT). The reference intervals established by this audit will assist other diagnostic laboratories using this platform.
