Rapid flow cytometric method for measuring lymphocyte subset activation

Maino, Vernon C.; Suni, Maria; Ruitenberg, Joyce J.

doi:10.1002/cyto.990200205

Cited by 192 publications

(144 citation statements)

References 22 publications

Supporting

Mentioning

130

Contrasting

Unclassified

Order By: Relevance

“…CD25 (IL-2Rα) is a marker expressed principally on activated T cells and Th1, Th2 and Th17 cytokines production induced by almost all stimuli was also a clear evidence of T cells activation. On the other hand, previous reports have shown overall agreement with CD25 and proliferation assays when a recall specific antigen was used [23,24]. In our model, CD3, CD4 and CD8 markers did not increase their expression when PBMCs were stimulated.…”

Section: Discussionsupporting

confidence: 83%

Specific Immune Response Induced by a Lactobacillus Associated with a Pneumococcal Antigen in an “in vitro” Human Cells Model

Vintiñi¹

2012

J Vaccines Vaccin

View full text Add to dashboard Cite

In this work we assessed the immune response induced by combinations of live (LcV) and heat-killed (LcM) Lactobacillus casei CRL 431 as adjuvants associated with pneumococcal protective A protein (PppA) in peripheral blood mononuclear cells (PBMCs). LcV, LcM and their combinations with PppA stimulated T, B and NK cells. Thus, all stimuli increased CD25 expression in T CD3 lymphocytes, highest activation being reached with the combinations of LcV or LcM with an antigen (PppA+LcV, PppA+LcM). Expression of CD19 B cells marker was significantly increased in almost all treatments compared with non-stimulated PBMCs, except for PppA. All treatments increased CD86 expression in the LT population, while in B cells only LPS, PppA+LcV and PppA+LcM increased it. NK cells were significantly increased by LPS (P<0.05), PppA+LcM (P<0.01) and PppA+LcV (P<0.01) compared to non-stimulated PBMCs. PppA+LcV and PppA+LcM increased CD56 expression in both NKT and NK cells, while LcM expanded NKT population. Cytokine pattern analysis showed that LcV and LcM stimulated Th, Th2 and Th17 cytokines and exerted an important adjuvant effect when associated with PppA. Correlation with previous results obtained in animal models when the same experimental vaccine was nasally administered is discussed. Human PBMCs would be useful to evaluate the immune response of mucosal vaccines containing lactic acid bacteria associated with a specific antigen.

show abstract

Section: Discussionsupporting

confidence: 83%

Specific Immune Response Induced by a Lactobacillus Associated with a Pneumococcal Antigen in an “in vitro” Human Cells Model

Vintiñi¹

2012

J Vaccines Vaccin

View full text Add to dashboard Cite

show abstract

“…Peptide-specific IFN-␥ production was assayed by intracellular staining (23) after stimulation of PBMC with 1 g/ml SL9 peptide or 10 g/ml NV9 peptide and 1 g/ml of brefeldin A (Golgi Plug; BD PharMingen). Positive controls used CD2/2R and CD28 Abs both at 10 g/ml (BD Biosciences) (24).…”

Section: Ctl Ifn-␥ (Ifn-␥ Production)mentioning

confidence: 99%

Epitope Escape Mutation and Decay of Human Immunodeficiency Virus Type 1-Specific CTL Responses

Jamieson

Yang

Hultin

et al. 2003

The Journal of Immunology

View full text Add to dashboard Cite

To investigate possible mechanisms behind HIV-1 escape from CTL, we performed detailed longitudinal analysis of Gag (SLYNTVATL)- and RT (ILKEPVHGV)-specific CTL responses and plasma epitope sequences in five individuals. Among those with CTL against consensus epitope sequences, epitope mutations developed over several years, invariably followed by decay of the CTL targeting the consensus epitopes. The maturation state of the CTL varied among individuals and appeared to affect the rate of epitope mutation and CTL decay, despite similar IFN-γ production. Escape mutations were oligoclonal, suggesting fitness constraints. The timing of escape indicated that the net selective advantage of escape mutants was slight, further underscoring the importance of understanding factors determining selective pressure and viral fitness in vivo. Our data show surprisingly consistent decay of CTL responses after epitope escape mutation and provide insight into potential mechanisms for both immune failure and shifting CTL specificities.

show abstract

“…Figure 2a shows the kinetics of appearance of CD69, a marker of activation [26] on CD3 cells from two individuals over a 24-h period. An initial rapid upregulation of this marker during the first 3 h was followed by a plateau during the remaining 21 h of the assay period.…”

Section: Kinetics Of Induction Of Il-2 and Ifn-g In Cd3 T Cells In CDmentioning

confidence: 99%

Intracellular cytokine expression in whole blood preparations from normals and patients with atopic dermatitis

Ferry

Antrobus

Hužička

et al. 1997

Clinical and Experimental Immunology

View full text Add to dashboard Cite

SUMMARYIn recent years, the importance of characterizing the role of cytokines in a wide range of clinical conditions has resulted in development of new methods to assess cytokine expression in clinical samples. The use of anti-cytokine MoAbs and flow cytometry to detect cytokines intracellularly at the single-cell level has the potential to quantify cytokine production in different diseases. For this technique to be useful in a clinical setting, rapid throughput of clinical samples and a cheap, reliable assay would be required, therefore the development of the above technique using unseparated whole blood samples would be advantageous. Using this technique, only one study to date (Maino et al., 1996) has used unseparated whole blood as the source of cells for detecting intracellular cytokines. In clinical practice, whole blood may be optimal, since this most closely approximates conditions in vivo: as no purification of blood mononuclear cells is required, very little blood is needed to detect a number of cytokines simultaneously in various lymphocyte subpopulations, and the assay can be applied to samples from infants and children. In this study we describe an intracellular cytokine assay using unseparated whole blood from normals. In activated CD8 ¹ T cells, IL-2 and interferon-gamma (IFN-g) were optimally induced after 10 h stimulation with phorbol 12-myristate acetate (PMA)/ionomycin, and in CD8 þ T cells IL-2 was optimally induced after 10 h and IFN-g after 6 h. The levels of IL-2 and IFN-g in CD8 þ and CD8 ¹ T cells in four healthy individuals were consistent on four occasions over a 3-month period. In a large group of 34 normal subjects, there was considerable heterogeneity in CD3/IL-2 þ (range 9 . 7-41 . 3) and CD3/IFN-g þ cells (10 . 1-44), expressed as a percentage of total lymphocytes. In patients with atopic dermatitis (n ¼ 5) there was a significantly decreased percentage of CD3 þ /CD8 þ peripheral blood T cells expressing IFN-g and an increased percentage of CD3 þ /CD8 ¹ T cells expressing IL-4 compared with non-atopic dermatitis controls (n ¼ 5). Possible applications of this technique are discussed.

show abstract

Rapid flow cytometric method for measuring lymphocyte subset activation

Cited by 192 publications

References 22 publications

Specific Immune Response Induced by a Lactobacillus Associated with a Pneumococcal Antigen in an “in vitro” Human Cells Model

Specific Immune Response Induced by a Lactobacillus Associated with a Pneumococcal Antigen in an “in vitro” Human Cells Model

Epitope Escape Mutation and Decay of Human Immunodeficiency Virus Type 1-Specific CTL Responses

Intracellular cytokine expression in whole blood preparations from normals and patients with atopic dermatitis

Contact Info

Product

Resources

About