HIV-Infected Spleens Present Altered Follicular Helper T Cell (Tfh) Subsets and Skewed B Cell Maturation
Fig 3
HIV infection severely impacts transcription profile of splenic Tfh and GCTfh cells.
GCTfh and Tfh were sorted using flow cytometry based on the expression of CXCR5 and PD-1 associated or not with CD57 marker respectively (S2 Fig). Sorted cells were used to perform high throughput gene expression measurement with real time PCR in a microfluidic dynamic array. After cleansing and normalization, consistent data were used for statistical analysis. We focused on genes specifically implicated in T cell functions (S1 Table). (A) 2D Principal component analysis (PCA) plot representation of gene expression profile of Tfh (triangle) and GCTfh (diamond) cells from HIV- ITP- (n = 4, empty symbols) and HIV+ ITP+ (n = 5, filled symbols) spleens. The projection of the data on the first and second principal components efficiently discriminates HIV+ samples, suggesting that HIV-infection deeply impacts gene expression profile of Tfh and GCTfh cells. (B) mRNA expression in Tfh and GCTfh sorted from splenocytes of uninfected (n = 4) and HIV+ (n = 5) individuals. The data are expressed in 40-Ct where Ct represents the threshold cycle number and 40 is chosen because the PCR run stops after 40 cycles. This value is directly correlated with the initial amount of RNA and therefore allows a quantitative comparison for 18 genes encoding cytokines/chemokines and proteins implicated in Tfh differentiation, costimulation, immune regulation and signal transduction. Statistics were obtained using the non parametric Mann-Whitney test *p<0.05, **p<0.005, ***p<0.001.
doi: https://doi.org/10.1371/journal.pone.0140978.g003