Article Title: What Is the most Important for Elite Control: Genetic Background of Patient, Genetic Background of Partner, both or neither? Description of Complete Natural History within a Couple of MSM
Figure Lengend Snippet: HIV infection and T-cell responses of PBMC from the case patient. (a) In vitro susceptibility to HIV infection in 2009. 9 CD4 + T-cells of the case patient or healthy donors (HD) were PHA-activated and infected in vitro with 100 ng p24 Gag of HIV YU2b (R5-tropic), HIV NL4–3 (× 4-tropic), and HIV KON a primary isolate of the same clade than CRF02_AG isolate. Viral replication was monitored by p24 Gag ELISA during 16 days post-infection (p.i.). (ni = non infected). (b) Ex vivo IFN-γ-ELISpot assay was performed in November 2009 using PBMC and lymph node (LN) cells loaded with 18 HIV-1 pools of 15-mers peptides overlapping by 11 amino acids. Eleven pools covered the three HIV type 1 Gag proteins: three pools for p17 Gag (1–55, 45–99, and 89–143), five pools for p24 Gag (133–187, 177–231, 221–275, 265–319, and 309–363), three pools for the small Gag proteins (Smp Gag) (p2/p7/p1/p6) (353–407, 397–451, and 441–512), four pools corresponding to poly-epitopic RT regions (293–352, 157–187, 177–216, and 4–52), and three pools corresponding to poly-epitopic Nef regions (181–206, 65–107, and 97–147). All results are expressed as specific IFN-γ-producing cells after subtracting the number of SFCs observed with cells alone, without stimulation. The IFN-γ producing HIV-specific CD8 + T cells were first investigated in an ELISpot assay detecting in 2009 significant responses to p17Gag (1–55) in PBMC and lymph node cells (110 SFC/10 6 cells) and to p24Gag (265–319) in PBMC alone (680 SFC/10 6 cells. (c) PBMC obtained in November 2009 were stimulated with the p17Gag 1 to 55 pool or with the p24 Gag 265 to 319 pool that elicited an IFN-γ-ELISpot production. The percentages of CD3 + CD8 + T cells producing IFN-γ, TNF-α or IL-2 were analyzed by intracellular cytokine staining and flow cytometry. The percentages of activated cells not subjected to peptide stimulation were subtracted. The immune-dominant CMV-derived HLA-B*07-restricted epitope (pp65 417 TPRVTGGGAM 426 ), and staphylococcal enterotoxin B (SEB) were used as positive controls. Cells alone served as negative control.
Article Snippet: Poly-functional NK was performed to simultaneously detect degranulation (anti-CD107a mAb, BD Biosciences) and cytokine production (intracellular expression IFN-γ (BD Biosciences) and TNF-α (E-Biosciences) ( ).
Techniques: Infection, In Vitro, Enzyme-linked Immunosorbent Assay, Ex Vivo, Enzyme-linked Immunospot, Staining, Flow Cytometry, Cytometry, Derivative Assay, Negative Control