γδ T Cell Characterization Services

γδ T cells are a minor T cell population in the peripheral blood (PB) from human adults, which are usually divided according to the type of Vδ chain they express at the T cell receptor (TCR). There are two major γδ T cells subsets: Vδ2 and Vδ1 T cells. Vδ2 chain associates with Vγ9 in most cases, defining a Vγ9Vδ2 Tc population that is unique to humans and other primates, whereas Vδ1 T cells use diverse Vγ regions. With proven experience and expertise in γδ T cells research, Creative Biolabs provides a comprehensive range of customized, high-quality contract R&D services in γδ T cells development and engineering, including γδ T cells characterization services.

γδ T Cell Phenotypic Characterization

Most peripheral blood γδ T cells are Vδ2+ and Vγ9+, and most of the remaining γδ T cells are Vδ1+. However, the percentages of Vδ1, Vδ2, and Vγ9 Tc are highly variable, varying from 3.5 to 65.7%, 15.7 to 96.0%, and 25.7 to 96.5%, respectively. In one-third of the individuals, the Vγ9Vδ2 subset accounted for <50% of the circulating γδ T cells. The median fraction of γδ T cells that were negative for Vδ1 and Vδ2 was 9.2%, ranging from 0.0% to 52.2%, corresponding to cells expressing Vδ chains that were not tested, such as Vδ3 and Vδ5.

Fig.1 Percentages of Vδ1+, Vδ2+, Vδ1-Vδ2-, Vγ9+, and Vγ9- T cells among peripheral blood γδ T cells. (Sónia, 2020)

Double-immunofluorescence staining by δCD8 and δCD4 showed considerable γ- and δ-positive cells exhibiting CD8-positive (δ+CD8+) and few δCD4 double-positive (δ+CD4+) existed. Detailed morphology of the γ+δ+ cells was detected at approximately 5 µm across, with round dense nuclei surrounded by a thin round ring of cytoplasm and numerous surface frills, as determined by transmission electron microscopy (TEM) and transmission electron microscopy (SEM). Flow cytometry (FCM) results showed the γ+δ+ T cell population consisted of 7.7, 9.3, 16.9, 18.3, and 20.5% of the total lymphocytes in the spleen, head kidney, skin, gills, and intestinal tissues, respectively. Results strongly indicated the existence of γδ T cell lineage in zebrafish, which exhibits phenotypic and morphological similarities to mammals.

Fig.2 Immunophenotype analysis of zebrafish γδ T cells by Abs against various surface molecules. (Feng, 2016)

γδ T Cell Functional Characterization

The phagocytic capacity of γδ T cells was initially examined through FCM. The γδ T cells were magnetically sorted from the selected peripheral blood lymphocyte (PBLs), spleens, and head kidney tissues and then examined to share a high degree of purity through FCM. This result showed that the γδ T cells could phagocytose substances in an actin polymerization-dependent manner. The enhanced amounts of Ag phagocytosis by γδ T cells were also determined based on the mean fluorescence intensity of Fluorescein isothiocyanate (FITC) and red fluorescence. Immunofluorescence confirmed that the red beads were indeed inside the cells. Zebrafish γδ T cells exhibit potent non-specific phagocytic abilities for both soluble and particulate antigens.

Fig.3 Phagocytic ability of zebrafish γδ T cells. (Feng, 2016)

Highly purified CD56+ and CD56- Vγ9Vδ2 T-cell subsets were isolated by FCM and their cytotoxic activities were examined. Both CD56+ and CD56- Vγ9Vδ2 T cells were able to kill fluA virus-infected cells. The cytotoxic ability of CD56+ Vγ9Vδ2 T cells against seasonal H1N1-infected MDMs was significantly higher than that of CD56- counterparts. Similar differences in the cytotoxic abilities of these 2 subsets against pdmH1N1, H9N2, or H5N1 virus-infected cells were also found. These results indicate that CD56+ Vγ9Vδ2 T-cell subsets have superior cytotoxic ability against fluA virus-infected target cells compared with their CD56- counterparts.

Fig.4 Cytotoxicity of CD56+ and CD56- γδ T-cell subsets against influenza A virus-infected monocyte-derived macrophages (MDMs). (G, 2012)

Our Services

• γδ T cells immunophenotype analysis by Abs
• γδ T cells morphology observation by TEM, SEM and FCM
• γδ T cells phagocytic ability tests
• Impedance-based label-free real-time γδ T cells cytotoxicity assay
• CAR transduction for engineered γδ T cell
• Drug-resistant engineered γδ T cells development services

Why Us?

• Having worked in the field of γδ T cell for many years.
• Covering complete delivery from design, implementation to a final report.
• One-stop customized service for γδ T cell.

With over 10 years of experience, Creative Biolabs is distinctly qualified to help you with your next γδ T cells project and affords the flexibility required to achieve the strictest timelines and goals. We understand that each project is unique and we draw from our vast experience to deliver you a custom solution that meets your needs. Contact us today to find out how we can help you.

References

1. Sónia, Fonseca.; et al. Human Peripheral Blood Gamma Delta T Cells: Report on a Series of Healthy Caucasian Portuguese Adults and Comprehensive Review of the Literature. 2020.
2. Feng, W.; et al. Characterization of γδ T Cells from Zebrafish Provides Insights into Their Important Role in Adaptive Humoral Immunity. Frontiers in Immunology. 2016, 7.
3. Qin, Y.; et al. Phenotypic and functional characterization of human γδ T cell subsets in response to influenza A viruses. International Journal of Infectious Diseases. 2012.