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. 2024 Oct 31;6(5):310-315.
doi: 10.35772/ghm.2024.01066.

Analysis of tumor infiltrating immune cells in Kaposi sarcoma lesions discovers shifts in macrophage populations

Takanobu Tagawa  1 , Guruswamy Mahesh  1 , Joseph M Ziegelbauer  1
Affiliations

Analysis of tumor infiltrating immune cells in Kaposi sarcoma lesions discovers shifts in macrophage populations

Takanobu Tagawa et al. Glob Health Med. .

Abstract

Limited information exists about the types of immune cells present in Kaposi sarcoma (KS) lesions, especially in KS in the gastrointestinal tract. Using previously reported RNA-sequencing results from Kaposi sarcoma lesions in skin and gastrointestinal tract with normal matched tissues from the same patients at the same time, we investigated changes in lymphocytes in these tissues. We employed a computational method that determines changes in cell type distributions using KS lesion transcriptome data compared to a reference set of RNA expression patterns of purified immune cells. Since secreted cytokines and chemokines from KSHV-infected cells may influence the microenvironment of Kaposi sarcoma lesions, we performed cytokine profiling of conditioned media from KSHV-infected primary human dermal lymphatic endothelial cells. We also measured how this conditioned media altered the differentiation of macrophages in cell culture assays. These results suggested that factors in conditioned media from KSHV-infected endothelial cells promoted differentiation of a promonocytic cell line to proinflammatory macrophages.

Keywords: KSHV; RNA-sequencing; cibersort.

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Conflict of interest statement

The authors have no conflicts of interest to disclose.

Figures

Figure 1.
Figure 1.
CIBERSORT analysis of KS lesions, cytokine secretion, and macrophage polarization. (A-B) Fold changes of different immune cell types in tumor vs normal samples, were estimated by CIBERSORT. Violin plots show the increased or decreased fold change (log2) of each cell type signature of the KS lesions as compared to normal skin. (C) Correlation analysis of changes in IFNG RNA expression (tumor vs. normal) and changes in M1 macrophage cell signatures (tumor vs. normal). (D) Proinflammatory cytokine levels in conditioned media of KSHV-infected LECs after 3 days were quantitated with an electrochemiluminescence multiplex immunoassay. Each line represents a separate biological replicate. Only cytokines with concentration > 5 pg/mL in any of samples are shown. (E) Macrophage polarization assays with conditioned media and a promonocytic cell line THP-1. THP-1 cells were treated with PMA and then incubated with conditioned media of KSHV-infected LECs (D) for 24 h. Transcript levels of M1 macrophage marker gene CXCL10 was quantitated with RT-qPCR and normalized to RPS13. Multiple paired t-test (two sided) was performed. *p < 0.05.
Figure 2.
Figure 2.
Correlation analysis of CIBERSORT values and KSHV infection. Normalized KSHV transcripts in the RNA-sequencing results were summarized and reported as total KSHV transcripts per million reads (KSHV TPM). Changes in CIBERSORT values were analyzed in combination with KSHV TPM values. Blue colors indicate strong positive correlations and red colors show strong negative correlations in all samples (A), only GI samples (B), and only skin samples (C).
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