CD32-RNA Co-localizes with HIV-RNA in CD3+ Cells Found within Gut Tissues from Viremic and ART-Suppressed Individuals

Joshua Jorge Vasquez, Brandon L. Aguilar-Rodriguez, Leonardo Rodriguez, Louise E. Hogan, Ma Somsouk, Joseph M. McCune, Steven G. Deeks, Zoltan G. Laszik, Peter W. Hunt, Timothy J. Henrich

Abstract


Background: Identifying biomarkers for cells harboring replication-competent HIV is a major research priority. Recently, there have been mixed reports addressing the possibility that CD32-expressing T-cells are enriched for HIV. There is growing evidence that CD32 expression increases with cellular activation that may be related to, but not necessarily specific for, infection with HIV.  However, the relationship of CD32 expression to HIV-infection in subtypes of tissue-resident leukocytes is unclear. 

 Methods: First, we used duplex chromogenic in situ hybridization to identify cells actively transcribing RNA for both CD32 and HIV on human gut tissues. Then we performed multiplexed immunofluorescence and in situ hybridization (mIFISH) on sections from the same tissues to determine the phenotype of individual cells co-expressing HIV-RNA and CD32-RNA. 

 Results: HIV-RNA+ cells were more abundant in tissues from viremic individuals compared to those on suppressive anti-retroviral therapy (ART). However, staining by both methods indicated that a higher proportion of HIV-RNA+ cells co-expressed CD32-RNA in ART-suppressed compared to viremic individuals. The majority of HIV-RNA+ cells were CD3+.

Conclusions: Our data suggest that the transcription of CD32-RNA is correlated with HIV transcriptional activity in CD3+ cells found within human gut tissue. Whether or not up-regulation of CD32-RNA is a direct result of HIV reactivation or more global T cell activation remains unclear.


Keywords


HIV; CD32; gut tissue; HIV reservoirs; HIV-RNA; in situ hybridization

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References


1. Churchill MJ, Deeks SG, Margolis DM, Siliciano RF, Swanstrom R. HIV reservoirs: what, where and how to target them. Nat Rev Microbiol. 2016;14(1):55-60. PubMed PMID: 26616417. doi: 10.1038/nrmicro.2015.5

2. Fletcher CV, Staskus K, Wietgrefe SW, Rothenberger M, Reilly C, Chipman JG, Beilman GJ, Khoruts A, Thorkelson A, Schmidt TE, Anderson J, Perkey K, Stevenson M, Perelson AS, Douek DC, Haase AT, Schacker TW. Persistent HIV-1 replication is associated with lower antiretroviral drug concentrations in lymphatic tissues. Proc Natl Acad Sci U S A. 2014;111(6):2307-12. PubMed PMID: 24469825. Pubmed Central PMCID: 3926074. doi: 10.1073/pnas.1318249111

3. Estes JD, Kityo C, Ssali F, Swainson L, Makamdop KN, Del Prete GQ, Deeks SG, Luciw PA, Chipman JG, Beilman GJ, Hoskuldsson T, Khoruts A, Anderson J, Deleage C, Jasurda J, Schmidt TE, Hafertepe M, Callisto SP, Pearson H, Reimann T, Schuster J, Schoephoerster J, Southern P, Perkey K, Shang L, Wietgrefe SW, Fletcher CV, Lifson JD, Douek DC, McCune JM, Haase AT, Schacker TW. Defining total-body AIDS-virus burden with implications for curative strategies. Nat Med. 2017;23(11):1271-6. PubMed PMID: 28967921. Pubmed Central PMCID: 5831193. doi: 10.1038/nm.4411

4. Pillai SK, Deeks SG. Signature of the Sleeper Cell: A Biomarker of HIV Latency Revealed. Trends Immunol. 2017;38(7):457-8. PubMed PMID: 28511815. Pubmed Central PMCID: 6091624. doi: 10.1016/j.it.2017.04.007

5. Descours B, Petitjean G, Lopez-Zaragoza JL, Bruel T, Raffel R, Psomas C, Reynes J, Lacabaratz C, Levy Y, Schwartz O, Lelievre JD, Benkirane M. CD32a is a marker of a CD4 T-cell HIV reservoir harbouring replication-competent proviruses. Nature. 2017;543(7646):564-7. PubMed PMID: 28297712. doi: 10.1038/nature21710

6. Badia R, Ballana E, Castellvi M, Garcia-Vidal E, Pujantell M, Clotet B, Prado JG, Puig J, Martinez MA, Riveira-Munoz E, Este JA. CD32 expression is associated to T-cell activation and is not a marker of the HIV-1 reservoir. Nat Commun. 2018;9(1):2739. PubMed PMID: 30013105. Pubmed Central PMCID: 6048139. doi: 10.1038/s41467-018-05157-w

7. Martin GE, Pace M, Thornhill JP, Phetsouphanh C, Meyerowitz J, Gossez M, Brown H, Olejniczak N, Lwanga J, Ramjee G, Kaleebu P, Porter K, Willberg CB, Klenerman P, Nwokolo N, Fox J, Fidler S, Frater J. CD32-Expressing CD4 T Cells Are Phenotypically Diverse and Can Contain Proviral HIV DNA. Front Immunol. 2018;9:928. PubMed PMID: 29780387. Pubmed Central PMCID: 5946760. doi: 10.3389/fimmu.2018.00928

8. Osuna CE, Lim SY, Kublin JL, Apps R, Chen E, Mota TM, Huang SH, Ren Y, Bachtel ND, Tsibris AM, Ackerman ME, Jones RB, Nixon DF, Whitney JB. Evidence that CD32a does not mark the HIV-1 latent reservoir. Nature. 2018;561(7723):E20-E8. PubMed PMID: 30232424. doi: 10.1038/s41586-018-0495-2

9. Bertagnolli LN, White JA, Simonetti FR, Beg SA, Lai J, Tomescu C, Murray AJ, Antar AAR, Zhang H, Margolick JB, Hoh R, Deeks SG, Tebas P, Montaner LJ, Siliciano RF, Laird GM, Siliciano JD. The role of CD32 during HIV-1 infection. Nature. 2018;561(7723):E17-E9. PubMed PMID: 30232425. Pubmed Central PMCID: 6442722. doi: 10.1038/s41586-018-0494-3

10. Perez L, Anderson J, Chipman J, Thorkelson A, Chun TW, Moir S, Haase AT, Douek DC, Schacker TW, Boritz EA. Conflicting evidence for HIV enrichment in CD32(+) CD4 T cells. Nature. 2018;561(7723):E9-E16. PubMed PMID: 30232423. Pubmed Central PMCID: 6410373. doi: 10.1038/s41586-018-0493-4

11. Descours B, Petitjean G, Benkirane M. Descours et al. reply. Nature. 2018;561(7723):E29. PubMed PMID: 30232426. doi: 10.1038/s41586-018-0496-1

12. Abdel-Mohsen M, Kuri-Cervantes L, Grau-Exposito J, Spivak AM, Nell RA, Tomescu C, Vadrevu SK, Giron LB, Serra-Peinado C, Genesca M, Castellvi J, Wu G, Del Rio Estrada PM, Gonzalez-Navarro M, Lynn K, King CT, Vemula S, Cox K, Wan Y, Li Q, Mounzer K, Kostman J, Frank I, Paiardini M, Hazuda D, Reyes-Teran G, Richman D, Howell B, Tebas P, Martinez-Picado J, Planelles V, Buzon MJ, Betts MR, Montaner LJ. CD32 is expressed on cells with transcriptionally active HIV but does not enrich for HIV DNA in resting T cells. Sci Transl Med. 2018;10(437). PubMed PMID: 29669853. Pubmed Central PMCID: 6282755. doi: 10.1126/scitranslmed.aar6759

13. Vasquez JJ, Hussien R, Aguilar-Rodriguez B, Junger H, Dobi D, Henrich TJ, Thanh C, Gibson E, Hogan LE, McCune J, Hunt PW, Stoddart CA, Laszik ZG. Elucidating the Burden of HIV in Tissues Using Multiplexed Immunofluorescence and In Situ Hybridization: Methods for the Single-Cell Phenotypic Characterization of Cells Harboring HIV In Situ. J Histochem Cytochem. 2018;66(6):427-46. PubMed PMID: 29462571. Pubmed Central PMCID: 5977441. doi: 10.1369/0022155418756848

14. Comber PG LV, Chien P. Interferon-gamma (IFN-gamma) and dexamethasone (DMS) modulate monocyte Fc-gamma receptor mRNA at the transcriptional level. A ASFJ. 1990.

15. Alevy YG, Tucker J, Naziruddin B, Mohanakumar T. CD32C (Fc gamma RIIC) mRNA expression and regulation. Mol Immunol. 1993;30(8):775-82. PubMed PMID: 8502244.

16. Alevy YG, Tucker J, Mohanakumar T. CD32A (Fc gamma RIIa) mRNA expression and regulation in blood monocytes and cell lines. Mol Immunol. 1992;29(11):1289-97. PubMed PMID: 1406718.

17. Sandilands GP, MacPherson SA, Burnett ER, Russell AJ, Downie I, MacSween RN. Differential expression of CD32 isoforms following alloactivation of human T cells. Immunology. 1997;91(2):204-11. PubMed PMID: 9227318. Pubmed Central PMCID: 1363848.

18. Comber PG, Lentz V, Schreiber AD. Modulation of the transcriptional rate of Fc gamma receptor mRNA in human mononuclear phagocytes. Cell Immunol. 1992;145(2):324-38. PubMed PMID: 1451181.

19. de la Salle C, Esposito-Farese ME, Bieber T, Moncuit J, Morales M, Wollenberg A, de la Salle H, Fridman WH, Cazenave JP, Teillaud JL, et al. Release of soluble Fc gamma RII/CD32 molecules by human Langerhans cells: a subtle balance between shedding and secretion? J Invest Dermatol. 1992;99(5):15S-7S. PubMed PMID: 1431201.

20. Sandilands GP, MacPherson SA, Burnett ER, Russell AJ, Downie I, MacSween RN. Differential expression of CD32 isoforms following alloactivation of human T cells. Immunology. 1997;91(2):204-11. PubMed PMID: 9227318. Pubmed Central PMCID: 1363848.

21. Wong JK, Yukl SA. Tissue reservoirs of HIV. Curr Opin HIV AIDS. 2016;11(4):362-70. PubMed PMID: 27259045. Pubmed Central PMCID: 4928570. doi: 10.1097/COH.0000000000000293

22. Banga R, Procopio FA, Noto A, Pollakis G, Cavassini M, Ohmiti K, Corpataux JM, de Leval L, Pantaleo G, Perreau M. PD-1(+) and follicular helper T cells are responsible for persistent HIV-1 transcription in treated aviremic individuals. Nat Med. 2016;22(7):754-61. PubMed PMID: 27239760. doi: 10.1038/nm.4113

23. Zandieh A, Izad M, Fakhri M, Amirifard H, Khazaeipour Z, Harirchian MH. Cytometric profiling in various clinical forms of multiple sclerosis with respect to CD21+, CD32+, and CD35+ B and T cells. Transl Neurodegener. 2013;2(1):14. PubMed PMID: 23819946. Pubmed Central PMCID: 3706361. doi: 10.1186/2047-9158-2-14

24. Engelhardt W, Matzke J, Schmidt RE. Activation-dependent expression of low affinity IgG receptors Fc gamma RII(CD32) and Fc gamma RIII(CD16) in subpopulations of human T lymphocytes. Immunobiology. 1995;192(5):297-320. PubMed PMID: 7649565.


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