Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022 Oct 9;14(10):2218.
doi: 10.3390/v14102218.

Macrophage Migration Inhibitory Factor (MIF) Promotes Increased Proportions of the Highly Permissive Th17-like Cell Profile during HIV Infection

César Trifone  1   2 , Lucía Baquero  2   3 , Alejandro Czernikier  1   2 , Paula Benencio  1   2 , Lin Leng  4 , Natalia Laufer  2   5 , María Florencia Quiroga  2   5 , Richard Bucala  4 , Yanina Ghiglione  1   2 , Gabriela Turk  2   5
Affiliations

Macrophage Migration Inhibitory Factor (MIF) Promotes Increased Proportions of the Highly Permissive Th17-like Cell Profile during HIV Infection

César Trifone et al. Viruses. .

Abstract

In this study, we evaluate the role of the MIF/CD74 axis in the functionality of CD4+ T lymphocytes (CD4TL) during HIV infection. MDMs from healthy donors were infected with a R5-tropic or Transmitted/Founder (T/F) HIV strain. At day 11 post-MDM infection, allogeneic co-cultures with uninfected CD4TLs plus MIF stimulus were performed. Cytokine production was evaluated by ELISA. MIF plasma levels of people with HIV (PWH) were evaluated by ELISA. The phenotype and infection rate of CD4TLs from PWH were analyzed after MIF stimulus. Intracellular cytokines and transcription factors were evaluated by flow cytometry. Data were analyzed by parametric or non-parametric methods. The MIF stimulation of HIV-infected MDMs induced an increased expression of IL-6, IL-1β and IL-8. In CD4TL/MDM co-cultures, the MIF treatment increased IL-17A/RORγt-expressing CD4TLs. Higher concentrations of IL-17A in supernatants were also observed. These results were recapitulated using transmitted/founder (T/F) HIV-1 strains. The MIF treatment appeared to affect memory CD4TLs more than naïve CD4TLs. MIF blocking showed a negative impact on IL17A+CD4TL proportions. Higher MIF concentrations in PWH-derived plasma were correlated with higher IL-17A+CD4TL percentages. Finally, MIF stimulation in PWH-derived PBMCs led to an increase in Th17-like population. MIF may contribute to viral pathogenesis by generating a microenvironment enriched in activating mediators and Th17-like CD4TLs, which are known to be highly susceptible to HIV-1 infection and relevant to viral persistence. These observations establish a basis for considering MIF as a possible therapeutic target.

Keywords: HIV; MDM/CD4TL co-culture; MIF; immune pathogenesis.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Activation status of HIV-infected MDMs after MIF stimulus. (A) Concentration of IL-6, IL-1β IL-10 and IL-8 in supernatants from HIV-infected MDMs treated (dark red lines) or not (grey lines) with MIF. (B) CD80, CD86 and HLA-DR surface expression on HIV-infected MDMs treated (dark red bars) or not (grey bars) with MIF after 72 and 120 h of stimulus. Data show the mean ± SD of three independent experiments each one performed in triplicate. Data analysis was performed by one-way ANOVA and Sidak’s post-test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.
Figure 2
Figure 2
CD4TL differentiation profile after five days of co-culture with HIV-infected MDMs treated with MIF. (A) Proportions of IL-17A-producing CD4TL (upper panel) and IFN-γ-producing CD4TL (lower panel); (B) Concentration of soluble IL-17A (upper panel) and IFN-γ (lower panel); (C) Proportions of RORγT-expressing CD4TLs (upper panel) and T-bet-expressing CD4TLs (lower panel) in MIF-treated and untreated co-cultures. (D) Expression of activation markers CD38 and HLA-DR on CD4TLs, CD80, CD86 and HLA-DR on MDMs and concentrations of IL-2, IL-6 and IL-10 in culture supernatants. Dark red bars: MIF-treated cultures. Grey bars: untreated cultures. Each point represents an independent experiment comprising triplicates. Bars indicate mean ± SD of all independent experiments. Data analysis was performed by Student’s t-test. * p < 0.05, ** p < 0.01.
Figure 3
Figure 3
HIV infection on CD4TL and viral production on infected co-cultures. (A) Proportion of p24+ events within CD4TL bulk population. (B) p24 viral protein concentration in culture supernatants. (C) HIV DNA pro-viral DNA copies per million of cells within CD4TL bulk population. (D) Proportion of p24+ events within Th17-like and Th1-like CD4 T-cell populations. Data represent the mean ± SD of three independents experiments each one performed in triplicate. Data analysis was performed with Student’s t-test (* p < 0.05).
Figure 4
Figure 4
Study of the mechanism involved in Th17-like profile promotion. IL-17A and IFNγ-expressing CD4TLs after conditioning with HIV-infected MIF-stimulated MDM-derived supernatant in parallel with (A) PHA stimulation, (B) CD3/CD28-engaging beads stimulation and (C) CD3/CD28-engaging antibodies stimulation. (D) Proportion of IL-17A-producing CD4TLs after MIF blocking in the context of an MIF-treated cell co-culture. From left to right: cell co-culture control with 25 ng/mL of MIF, immunoglobulin isotype control, MIF-neutralizing monoclonal antibody, MIF chemical antagonist (#98), IL-6 and IL-1β neutralizing monoclonal antibodies. Each point represents an independent experiment comprising triplicates. Bars indicate the mean ± SD of all independent experiments each one performed in triplicate. Data analysis was performed by a one-way ANOVA and Tukey’s post-test (excluding the positive control). * p < 0.05, ** p < 0.01.
Figure 5
Figure 5
Transmitted/founder (T/F) HIV-strain MDM infection of co-culture resembles Th17-like profiling. (A) Proportion of IL-17A-expressing CD4TLs on MIF-treated co-cultures with T/F HIV-strain-infected MDMs. (B) Soluble IL-17A concentration in culture supernatants on MIF-treated co-cultures with T/F HIV-strain-infected MDMs. Data show the mean of two independents experiments with five different T/F HIV strains each one performed in triplicate. Data analysis was performed with a Student’s t-test (* p < 0.05).
Figure 6
Figure 6
Study of the response of naïve and memory (CD4TL populations to MIF stimulus. Proportion of IL-17A expressing CD4TL in cell co-cultures performed with BAL R5-infected MDMs and: (A) sorted naïve CD4TLs (CD45RA+CCR7high) and (B) sorted memory CD4TLs (CD45RO+CCR7−). Individual experiments mean values (upper panel), all replicates (below panel). Data represent the mean ± SD of three independent experiments each one performed in triplicate. Data analysis was performed with a Student’s t-test (* p < 0.05).
Figure 7
Figure 7
Association between MIF concentration in plasma and the proportion of IL-17A-expressing CD4TL in PBMCs, both derived from PWH. (A) Ex vivo IL-17A-expressing CD4TL percentage in PBMC from PWH segregated by low (<100 ng/mL) or high (>100 ng/mL) MIF plasma concentration. Data represent the median ± IQR25-75 of 15 donors. Data analysis was performed with a Mann–Whitney test. * p < 0.05. (B) Concentration of soluble IL-17A in MIF-stimulated co-cultures after five days of treatment with 100 ng/mL of MIF. MDM infection was performed with T/F HIV strains. Data show the average of means of at least three independent experiments with four different T/F HIV strains and the BAL R5 strain as a control, each one performed in triplicate, relative to the untreated control. Data analysis was performed with a Mann–Whitney test ** p < 0.01. (C) Concentration of soluble IFNγ in MIF-treated co-cultures after five days of treatment with 100 ng/mL of MIF with T/F HIV-strain-infected MDMs. Data show the average of means of at least three independent experiments with four different T/F HIV strains and the BAL R5 strain as a control, each one performed in triplicate, relative to the untreated control. Data analysis was performed with a Student’s t-test.
Figure 8
Figure 8
Evaluation of PBMCs from PWH response to MIF stimulus in an ex vivo model. (A) Ex vivo HIV-infection percentage in IL-17A- and IFNγ-expressing CD4TLs derived from PBMCs of PWH. (B) IFNγ intracellular expression on CD4TLs after 24 h of treatment with 100 ng/mL of MIF. (C) IL-17A intracellular expression on CD4TLs after 24 h of treatment with 100 ng/mL of MIF. Both experiments were performed on PBMCs and the results were standardized to the positive control (PBMCs treated with αCD3/αCD28-binding antibodies). Data represent the median ± IQR25-75 of 15 independent donors, each one with one replicate. Data analysis was performed with a Student’s t-test. ** p < 0.01, *** p < 0.001.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Maartens G., Celum C., Lewin S.R. HIV infection: Epidemiology, pathogenesis, treatment, and prevention. Lancet. 2014;384:258–271. doi: 10.1016/S0140-6736(14)60164-1. - DOI - PubMed
    1. Okoli C., Van de Velde N., Richman B., Allan B., Castellanos E., Young B., Brough G., Eremin A., Corbelli G.M., Mc Britton M., et al. Undetectable equals untransmittable (U = U): Awareness and associations with health outcomes among people living with HIV in 25 countries. Sex. Transm. Infect. 2021;97:18–26. doi: 10.1136/sextrans-2020-054551. - DOI - PMC - PubMed
    1. Deeks S.G., Tracy R., Douek D.C. Systemic effects of inflammation on health during chronic HIV infection. Immunity. 2013;39:633–645. doi: 10.1016/j.immuni.2013.10.001. - DOI - PMC - PubMed
    1. Lv T., Cao W., Li T. HIV-Related Immune Activation and Inflammation: Current Understanding and Strategies. J. Immunol. Res. 2021;2021:7316456. doi: 10.1155/2021/7316456. - DOI - PMC - PubMed
    1. Fromentin R., Chomont N. HIV persistence in subsets of CD4+ T cells: 50 shades of reservoirs. Semin. Immunol. 2021;51:101438. doi: 10.1016/j.smim.2020.101438. - DOI - PMC - PubMed

Publication types

MeSH terms