Complement opsonization of HIV affects primary infection of human colorectal mucosa and subsequent activation of T cells
- Division of Molecular Medicine and Virology, Department of Clinical and Experimental Medicine, Linköping University, Sweden
- Division of Surgery, Orthopedics and Oncology, Linköping University, Sweden
- Center of Excellence for Research in AIDS (CERiA), University of Malaya, Lembah Pantai, Malaysia
- Division of Infection Biology and Medical Microbiology, Department of Life Sciences, Central University of Tamil Nadu, India
- Department of Clinical Immunology and Transfusion Medicine and Department of Clinical and Experimental Medicine, Linköping University, Sweden
Figures
Figure 1 with 3 supplements
HIV-induced activation of innate and inflammatory pathways in colorectal mucosa with high level of antiviral factors in F-HIV.
The colorectal tissue biopsies were exposed to HIV-1BaL (250 ng/mL), either free (F-HIV), complement-opsonized (C-HIV), or virions opsonized by a cocktail of complement and antibodies (CI-HIV) or …
Figure 1—figure supplement 1
Histological images of colonic mucosa.
Representative histological images from three different donors of hematoxylin-eosin staining of sections from colonic mucosa used in the study. Original magnification 50x.
Figure 1—figure supplement 2
Complement and IgG opsonization of HIV-1.
(A) The generation of iC3b was assessed in serum incubated without HIV-1 (MOCK) or in serum incubated with HIV-1 for 1 hr at 37°C. HIV-1 alone without serum was used as a control. The levels of iC3b …
Figure 1—figure supplement 3
Complement opsonization elevated the antiviral and HIV regulatory factors in colorectal tissue.
Mucosal immune cells isolated from the colorectal tissue biopsies by enzymatic digestion were exposed to HIV-1BaL, either free (F-HIV), complement opsonized (C-HIV), or virions opsonized by a …
Figure 2 with 1 supplement
Complement opsonization elevated the levels of HIV-infected colorectal mucosal immune cells.
The colorectal tissue biopsies were exposed to HIV-1BaL (250 ng/mL), either free (F-HIV), complement-opsonized (C-HIV), or virions opsonized by a cocktail of complement and antibodies (CI-HIV) or …
Figure 2—figure supplement 1
Representative Zebra plots with gating for HIV-1 p24+ Immune cells.
Mucosal immune cells isolated from the colorectal tissue biopsies by enzymatic digestion were exposed to HIV-1BaL, either free (F-HIV), complement-opsonized (C-HIV), or virions opsonized by a …
Figure 3 with 1 supplement
Alteration in mucosal CD4+ and CD8+ T cell memory cell after HIV-1 exposure.
Mucosal immune cells isolated from the colorectal tissue biopsies by enzymatic digestion were exposed to HIV-1BaL (250 ng/mL), either free (F-HIV), complement-opsonized (C-HIV), or virions opsonized …
Figure 3—figure supplement 1
The gating strategy for the CD4+ and CD8+ T cell subsets and the viability of the CD3+ T cells.
(A-B) Strategy for gating of T cell subsets and T cell viability. Mucosal immune cells isolated from the colorectal tissue biopsies by enzymatic digestion were exposed to HIV-1BaL, either free …
Figure 4 with 1 supplement
HIV-1 exposure alters the T cell expression of transcriptional factors and chemokine receptors.
Mucosal immune cells isolated from the colorectal tissue biopsies by enzymatic digestion were exposed to HIV-1BaL (250 ng/mL), either free (F-HIV), complement-opsonized (C-HIV), or virions opsonized …
Figure 4—figure supplement 1
Exposure to HIV modulated the transcription factor and cytokines associated with different T-cell subsets in the isolated mucosal immune cells.
Mucosal immune cells isolated from the colorectal tissue biopsies by enzymatic digestion were exposed to HIV-1BaL, either free (F-HIV), complement-opsonized (C-HIV), or virions opsonized by a …
Figure 5 with 1 supplement
HIV-1 exposure alters the levels of negative immune checkpoint factors on the mucosal T cells.
Mucosal immune cells isolated from the colorectal tissue biopsies by enzymatic digestion were exposed to HIV-1BaL (250 ng/mL), either free (F-HIV), complement-opsonized (C-HIV), or virions opsonized …
Figure 5—figure supplement 1
Representative Zebra plots for immune checkpoint factors PD1 and LAG3 on mucosal T cells.
Mucosal immune cells isolated from the colorectal tissue biopsies by enzymatic digestion were exposed to HIV-1BaL for 96 hr, either free (F-HIV), complement-opsonized (C-HIV), or virions opsonized …
Figure 6
HIV exposure modulated transcriptional regulation of mucosal T cells.
Mucosal immune cells isolated from the colorectal tissue biopsies by enzymatic digestion were exposed to HIV-1BaL (250 ng/mL), either free (F-HIV), complement-opsonized (C-HIV), or virions opsonized …
Figure 7
HIV exposure alters the expression of perforin and granzyme in mucosal cells.
Mucosal immune cells isolated from the colorectal tissue biopsies by enzymatic digestion were exposed to HIV-1BaL (250 ng/mL), either free (F-HIV), complement-opsonized (C-HIV), or virions opsonized …
Figure 8
Complement-opsonized HIV gave rise to a delayed inflammation in the mucosal immune cells.
Mucosal immune cells isolated from the colorectal tissue biopsies by enzymatic digestion were exposed to HIV-1BaL (250 ng/mL), either free (F-HIV), complement-opsonized (C-HIV), or virions opsonized …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Biological sample (human) | Colorectal Tissue Biopsies | Surgical Department of Linköping University Hospital | The tissue pieces were either cut into small explants or punched into biopsies | |
| Antibody | anti-human CD3-PerCP/AmCyan(Mouse monoclonal) | BD Biosciences | Cat#: 552851/339186 | FACS (1 ul per test) |
| anti-human CD4-PE/BV421(Mouse monoclonal) | BD Biosciences | Cat#: 555347/562424 | FACS (1 ul per test) | |
| anti-human CD8-PerCPCy5.5(Mouse monoclonal) | BD Biosciences | Cat#: 560662 | FACS (1 ul per test) | |
| anti-human CD1a-APC(Mouse monoclonal) | BD Biosciences | Cat#: 559775 | FACS (1 ul per test) | |
| anti-human CD163-APCCy7(Mouse monoclonal) | BioLegend | Cat#: 50-850-648 | FACS (1 ul per test) | |
| anti-human CD45RA-FITC(Mouse monoclonal) | BD Biosciences | Cat#: 555488 | FACS (1 ul per test) | |
| anti-human CD45RO-FITC(Mouse monoclonal) | BD Biosciences | Cat#: 555492 | FACS (1 ul per test) | |
| anti-human CD38-PerCP-Cy5.5(Mouse monoclonal) | BD Biosciences | Cat#: 551400 | FACS (1 ul per test) | |
| anti-human CD160-PE(Mouse monoclonal) | BD Biosciences | Cat#: 562118 | FACS (1 ul per test) | |
| anti-human PD-1-BV421(Mouse monoclonal) | BioLegend | Cat#: 329920 | FACS (1 ul per test) | |
| anti-human TIM3-PE(Mouse monoclonal) | BioLegend | Cat#: 345006 | FACS (1 ul per test) | |
| anti-human LAG3-FITC(Mouse monoclonal) | BioLegend | Cat#: 369308 | FACS (1 ul per test) | |
| anti-human CXCR3-PE(Mouse monoclonal) | BioLegend | Cat#: 353706 | FACS (1 ul per test) | |
| anti-human CCR4-AF647(Mouse monoclonal) | BioLegend | Cat#: 359404 | FACS (1 ul per test) | |
| anti-human CCR6-PECy7(Mouse monoclonal) | BD Biosciences | Cat#: 560620 | FACS (1 ul per test) | |
| anti-human CCR7-PerCPCy5.5(Mouse monoclonal) | BioLegend | Cat#: 353220 | FACS (1 ul per test) | |
| anti-Tbet-FITC/PECy7(Mouse monoclonal) | BioLegend | Cat#: 644812/644824 | FACS (1 ul per test) | |
| anti-human EOMES-eFluor660(Mouse monoclonal) | eBioscience | Cat#: 50-4877-41 | FACS (1 ul per test) | |
| anti-human RORγt-PE(Mouse monoclonal) | BD Biosciences | Cat#: 563081 | FACS (1 ul per test) | |
| anti-GATA3-PECy7(Mouse monoclonal) | BD Biosciences | Cat#: 560405 | FACS (1 ul per test) | |
| anti-human FoxP3-AF647(Mouse monoclonal) | BD Biosciences | Cat#: 560045 | FACS (1 ul per test) | |
| anti-human Perforin-Pacific Blue(Mouse monoclonal) | BioLegend | Cat#: 308118 | FACS (1 ul per test) | |
| anti-human Granzyme B-AF647(Mouse monoclonal) | BD Biosciences | Cat#: 560212 | FACS (1 ul per test) | |
| Anti-human HIV-1 Core Antigen-FITC, KC57 (Mouse monoclonal) | Beckman Coulter | Cat#: 6604665 | FACS (1 ul per test) | |
| Anti-human IgG1 Mouse-FITC Isotype Control | Beckman Coulter | Cat#: IM0639U | FACS (1 ul per test) | |
| Zombie NIR Fixable Viability Kit | BioLegend | Cat#: 423106 | FACS (diluted as manufacturer’s protocol) | |
| Sequence-based reagent | β-Actin_F | Eurofins Genomics/CyberGene AB | PCR primers | |
| β-Actin_R | Eurofins Genomics/CyberGene AB | PCR primers | ||
| GADPH_F | Eurofins Genomics/CyberGene AB | PCR primers | ||
| GADPH_R | Eurofins Genomics/CyberGene AB | PCR primers | ||
| Commercial assay or kit | Human Inflammatory Cytokine CBA Kit | BD Biosciences | 551811 | |
| Human IFNγ CBA Flex Set | BD Biosciences | 558269 | ||
| Human IP-10 CBA Flex Set | BD Biosciences | 558280 | ||
| Human MIP-1α CBA Flex Set | BD Biosciences | 558325 | ||
| Human IL-17A Enhanced Sensitivity CBA Flex Set | BD Biosciences | 562143 | ||
| Software, algorithm | CFX Manager 3.1 | Bio-Rad | ||
| Ingenuity Pathway Analysis (IPA) | Qiagen | |||
| FlowJo | Treestar | |||
| FCAP Array | BD Biosciences | |||
| GraphPad Prism 5 | GraphPad Software |
