Modulation of HIVGP120 Antigen-Specific Immune Responses In Vivo by Δ9-Tetrahydrocannabinol

Abstract

Approximately 25 % of HIV patients use marijuana for its putative therapeutic benefit; however, it is unknown how cannabinoids affect the immune status of HIV patients. Previously, a surrogate in vitro mouse model was established, which induced CD8+ T cell proliferation and gp120-specific IFNγ production. ∆9-Tetrahydrocannabinol (THC), the predominant psychoactive compound in marijuana, suppressed or enhanced the responses depending on the magnitude of cellular activation. The purpose of the current study was to investigate whether THC produced similar effects in vivo and therefore a mouse model to induce HIVgp120-specific immune responses was established. A gp120-expressing plasmid, pVRCgp120, or a vector plasmid, pVRC2000, was injected intramuscularly into mice, which were also dosed with THC orally. The gp120-specific IFNγ and IL-2 responses were detected when splenocytes were restimulated with gp120-derived peptide 81 (IIGDIRQAHCNISRA), which was identified as being immunodominant. Various cellular populations were activated in response to pVRCgp120 stimulation followed by peptide restimulation, as evidenced by increased expression levels of activation markers (e.g., CD69, CD80, and major histocompatibility complex II [MHC II]). The IFNγ response and cellular activation were enhanced by THC in C57Bl/6 wild type (WT) mice but suppressed or not affected by THC in cannabinoid receptor 1 (CB1) and 2 (CB2) knockout (CB1 −/−CB2 −/−) mice. Furthermore, CB1 −/−CB2 −/− mice exhibited augmented IFNγ production when compared to WT mice in the absence of THC. Collectively, our findings demonstrate that under certain conditions, THC enhances HIV antigen-specific immune responses, which occurs through CB1/CB2-dependent and -independent mechanisms.

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Abbreviations

AC:

Adenylyl cyclase

AP-1:

Activator protein-1

APC:

Antigen presenting cell

BCS:

Bovine calf serum

cAMP:

Cyclic adenosine 3′,5′-monophosphate

CB1 :

Cannabinoid receptor 1

CB2 :

Cannabinoid receptor 2

CB1 −/−CB2 −/− :

CB1 and CB2 null

CO:

Corn oil

CTL:

Cytotoxic T lymphocytes

DC:

Dendritic cell

DMSO:

Dimethyl sulfoxide

FcRs:

Fragment cystallizable receptors

GPCR:

G protein-coupled receptor

i.m.:

Intramuscular

KV :

Voltage-gated potassium channels

MCP-1:

Monocyte chemotactic protein-1

MFI:

Mean fluorescence intensity

NA:

Naïve

NFAT:

Nuclear factor of activated T cells

NF-κB:

Nuclear factor kappa-light-chain-enhancer of activated B cells

NT:

No treatment

PBS:

Phosphate buffered saline

SIV:

Simian immunodeficiency virus

THC:

Δ9-tetrahydrocannabinol

TRP:

Transient receptor potential

VH:

Vehicle

WT:

Wild type

References

  1. Abrams DI, Hilton JF, Leiser RJ, Shade SB, Elbeik TA, Aweeka FT, Benowitz NL, Bredt BM, Kosel B, Aberg JA, Deeks SG, Mitchell TF, Mulligan K, Bacchetti P, McCune JM, Schambelan M (2003) Short-term effects of cannabinoids in patients with HIV-1 infection: a randomized, placebo-controlled clinical trial. Ann Intern Med 139:258–266

    CAS  PubMed  Article  Google Scholar 

  2. Azorlosa JL, Heishman SJ, Stitzer ML, Mahaffey JM (1992) Marijuana smoking: effect of varying delta 9-tetrahydrocannabinol content and number of puffs. J Pharmacol Exp Ther 261:114–122

    CAS  PubMed  Google Scholar 

  3. Barouch DH, Santra S, Tenner-Racz K, Racz P, Kuroda MJ, Schmitz JE, Jackson SS, Lifton MA, Freed DC, Perry HC, Davies ME, Shiver JW, Letvin NL (2002) Potent CD4+ T cell responses elicited by a bicistronic HIV-1 DNA vaccine expressing gp120 and GM-CSF. J Immunol 168:562–568

    CAS  PubMed  Article  Google Scholar 

  4. Billaut-Mulot O, Idziorek T, Loyens M, Capron A, Bahr GM (2001) Modulation of cellular and humoral immune responses to a multiepitopic HIV-1 DNA vaccine by interleukin-18 DNA immunization/viral protein boost. Vaccine 19:2803–2811

    CAS  PubMed  Article  Google Scholar 

  5. Billaut-Mulot O, Idziorek T, Ban E, Kremer L, Dupre L, Loyens M, Riveau G, Locht C, Capron A, Bahr GM (2000) Interleukin-18 modulates immune responses induced by HIV-1 Nef DNA prime/protein boost vaccine. Vaccine 19:95–102

    CAS  PubMed  Article  Google Scholar 

  6. Bonhaus DW, Chang LK, Kwan J, Martin GR (1998) Dual activation and inhibition of adenylyl cyclase by cannabinoid receptor agonists: evidence for agonist-specific trafficking of intracellular responses. J Pharmacol Exp Ther 287:884–888

    CAS  PubMed  Google Scholar 

  7. Brown SA, Stambas J, Zhan X, Slobod KS, Coleclough C, Zirkel A, Surman S, White SW, Doherty PC, Hurwitz JL (2003) Clustering of Th cell epitopes on exposed regions of HIV envelope despite defects in antibody activity. J Immunol 171:4140–4148

    CAS  PubMed  Article  Google Scholar 

  8. Buchweitz JP, Karmaus PW, Harkema JR, Williams KJ, Kaminski NE (2007) Modulation of airway responses to influenza A/PR/8/34 by Delta9-tetrahydrocannabinol in C57BL/6 mice. J Pharmacol Exp Ther 323:675–683

    CAS  PubMed  Article  Google Scholar 

  9. Buchweitz JP, Karmaus PW, Williams KJ, Harkema JR, Kaminski NE (2008) Targeted deletion of cannabinoid receptors CB1 and CB2 produced enhanced inflammatory responses to influenza A/PR/8/34 in the absence and presence of Delta9-tetrahydrocannabinol. J Leukoc Biol 83:785–796

    CAS  PubMed  Article  Google Scholar 

  10. Busch L, Sterin-Borda L, Borda E (2004) Expression and biological effects of CB1 cannabinoid receptor in rat parotid gland. Biochem Pharmacol 68:1767–1774

    CAS  PubMed  Article  Google Scholar 

  11. Cabral GA, Mishkin EM, Marciano-Cabral F, Coleman P, Harris L, Munson AE (1986) Effect of delta 9-tetrahydrocannabinol on herpes simplex virus type 2 vaginal infection in the guinea pig. Proc Soc Exp Biol Med 182:181–186

    CAS  PubMed  Article  Google Scholar 

  12. Calandra B, Portier M, Kerneis A, Delpech M, Carillon C, Le Fur G, Ferrara P, Shire D (1999) Dual intracellular signaling pathways mediated by the human cannabinoid CB1 receptor. Eur J Pharmacol 374:445–455

    CAS  PubMed  Article  Google Scholar 

  13. Carbonetti NH, Tuskan RG, Lewis GK (2001) Stimulation of HIV gp120-specific cytolytic T lymphocyte responses in vitro and in vivo using a detoxified pertussis toxin vector. AIDS Res Hum Retrovir 17:819–827

    CAS  PubMed  Article  Google Scholar 

  14. Chandy KG, Wulff H, Beeton C, Pennington M, Gutman GA, Cahalan MD (2004) K+ channels as targets for specific immunomodulation. Trends Pharmacol Sci 25:280–289

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  15. Chen W, Kaplan BL, Pike ST, Topper LA, Lichorobiec NR, Simmons SO, Ramabhadran R, Kaminski NE (2012) Magnitude of stimulation dictates the cannabinoid-mediated differential T cell response to HIVgp120. J Leukoc Biol 92:1093–1102

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  16. Donnelly JJ, Ulmer JB, Shiver JW, Liu MA (1997) DNA vaccines. Annu Rev Immunol 15:617–648

    CAS  PubMed  Article  Google Scholar 

  17. Felder CC, Joyce KE, Briley EM, Mansouri J, Mackie K, Blond O, Lai Y, Ma AL, Mitchell RL (1995) Comparison of the pharmacology and signal transduction of the human cannabinoid CB1 and CB2 receptors. Mol Pharmacol 48:443–450

    CAS  PubMed  Google Scholar 

  18. Fischer HG, Eder C (1995) Voltage-gated K+ currents of mouse dendritic cells. FEBS Lett 373:127–130

    CAS  PubMed  Article  Google Scholar 

  19. Gaoni Y, Mechoulam R (1964) Isolation, structure, and partial synthesis of an active constituent of hashish. J Am Chem Soc 86:1646–1647

    CAS  Article  Google Scholar 

  20. Hai T, Curran T (1991) Cross-family dimerization of transcription factors Fos/Jun and ATF/CREB alters DNA binding specificity. Proc Natl Acad Sci U S A 88:3720–3724

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  21. Hovav AH, Santosuosso M, Bivas-Benita M, Plair A, Cheng A, Elnekave M, Righi E, Chen T, Kashiwagi S, Panas MW, Xiang SH, Furmanov K, Letvin NL, Poznansky MC (2009) X4 human immunodeficiency virus type 1 gp120 down-modulates expression and immunogenicity of codelivered antigens. J Virol 83:10941–10950

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  22. Howlett AC, Barth F, Bonner TI, Cabral G, Casellas P, Devane WA, Felder CC, Herkenham M, Mackie K, Martin BR, Mechoulam R, Pertwee RG (2002) International union of pharmacology. XXVII. Classification of cannabinoid receptors. Pharmacol Rev 54:161–202

    CAS  PubMed  Article  Google Scholar 

  23. Huestis MA, Gorelick DA, Heishman SJ, Preston KL, Nelson RA, Moolchan ET, Frank RA (2001) Blockade of effects of smoked marijuana by the CB1-selective cannabinoid receptor antagonist SR141716. Arch Gen Psychiatry 58:322–328

    CAS  PubMed  Article  Google Scholar 

  24. Jan TR, Kaminski NE (2001) Role of mitogen-activated protein kinases in the differential regulation of interleukin-2 by cannabinol. J Leukoc Biol 69:841–849

    CAS  PubMed  Google Scholar 

  25. Jan TR, Rao GK, Kaminski NE (2002) Cannabinol enhancement of interleukin-2 (IL-2) expression by T cells is associated with an increase in IL-2 distal nuclear factor of activated T cell activity. Mol Pharmacol 61:446–454

    CAS  PubMed  Article  Google Scholar 

  26. Jan TR, Farraj AK, Harkema JR, Kaminski NE (2003) Attenuation of the ovalbumin-induced allergic airway response by cannabinoid treatment in A/J mice. Toxicol Appl Pharmacol 188:24–35

    CAS  PubMed  Article  Google Scholar 

  27. Kaplan BL, Rockwell CE, Kaminski NE (2003) Evidence for cannabinoid receptor-dependent and -independent mechanisms of action in leukocytes. J Pharmacol Exp Ther 306:1077–1085

    CAS  PubMed  Article  Google Scholar 

  28. Kara CJ, Liou HC, Ivashkiv LB, Glimcher LH (1990) A cDNA for a human cyclic AMP response element-binding protein which is distinct from CREB and expressed preferentially in brain. Mol Cell Biol 10:1347–1357

    CAS  PubMed Central  PubMed  Google Scholar 

  29. Karmaus PW, Chen W, Crawford R, Kaplan BL, Kaminski NE (2013) Delta9-tetrahydrocannabinol impairs the inflammatory response to influenza infection: role of antigen-presenting cells and the cannabinoid receptors 1 and 2. Toxicol Sci 131:419–433

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  30. Karsak M, Gaffal E, Date R, Wang-Eckhardt L, Rehnelt J, Petrosino S, Starowicz K, Steuder R, Schlicker E, Cravatt B, Mechoulam R, Buettner R, Werner S, Di Marzo V, Tuting T, Zimmer A (2007) Attenuation of allergic contact dermatitis through the endocannabinoid system. Science 316:1494–1497

    CAS  PubMed  Article  Google Scholar 

  31. Klein TW, Newton CA, Nakachi N, Friedman H (2000) Delta 9-tetrahydrocannabinol treatment suppresses immunity and early IFN-gamma, IL-12, and IL-12 receptor beta 2 responses to Legionella pneumophila infection. J Immunol 164:6461–6466

    CAS  PubMed  Article  Google Scholar 

  32. Korber BT, Letvin NL, Haynes BF (2009) T-cell vaccine strategies for human immunodeficiency virus, the virus with a thousand faces. J Virol 83:8300–8314

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  33. Koup RA, Safrit JT, Cao Y, Andrews CA, McLeod G, Borkowsky W, Farthing C, Ho DD (1994) Temporal association of cellular immune responses with the initial control of viremia in primary human immunodeficiency virus type 1 syndrome. J Virol 68:4650–4655

    CAS  PubMed Central  PubMed  Google Scholar 

  34. Lee CY, Wey SP, Liao MH, Hsu WL, Wu HY, Jan TR (2008) A comparative study on cannabidiol-induced apoptosis in murine thymocytes and EL-4 thymoma cells. Int Immunopharmacol 8:732–740

    CAS  PubMed  Article  Google Scholar 

  35. Lu T, Newton C, Perkins I, Friedman H, Klein TW (2006) Role of cannabinoid receptors in Delta-9-tetrahydrocannabinol suppression of IL-12p40 in mouse bone marrow-derived dendritic cells infected with Legionella pneumophila. Eur J Pharmacol 532:170–177

    CAS  PubMed  Article  Google Scholar 

  36. Maneuf YP, Brotchie JM (1997) Paradoxical action of the cannabinoid WIN 55,212-2 in stimulated and basal cyclic AMP accumulation in rat globus pallidus slices. Br J Pharmacol 120:1397–1398

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  37. Matsuda LA, Lolait SJ, Brownstein MJ, Young AC, Bonner TI (1990) Structure of a cannabinoid receptor and functional expression of the cloned cDNA. Nature 346:561–564

    CAS  PubMed  Article  Google Scholar 

  38. Mirano-Bascos D, Tary-Lehmann M, Landry SJ (2008) Antigen structure influences helper T-cell epitope dominance in the human immune response to HIV envelope glycoprotein gp120. Eur J Immunol 38:1231–1237

    CAS  PubMed  Article  Google Scholar 

  39. Mo Y, Quanquin NM, Vecino WH, Ranganathan UD, Tesfa L, Bourn W, Derbyshire KM, Letvin NL, Jacobs WR Jr, Fennelly GJ (2007) Genetic alteration of mycobacterium smegmatis to improve mycobacterium-mediated transfer of plasmid DNA into mammalian cells and DNA immunization. Infect Immun 75:4804–4816

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  40. Molina PE, Winsauer P, Zhang P, Walker E, Birke L, Amedee A, Stouwe CV, Troxclair D, McGoey R, Varner K, Byerley L, Lamotte L (2010) Cannabinoid administration attenuates the progression of simian immunodeficiency virus. AIDS Res Hum Retrovir 27:585–592

    PubMed  Article  Google Scholar 

  41. Munro S, Thomas KL, Abu-Shaar M (1993) Molecular characterization of a peripheral receptor for cannabinoids. Nature 365:61–65

    CAS  PubMed  Article  Google Scholar 

  42. Overbaugh J, Morris L (2012) The Antibody Response against HIV-1. Cold Spring Harbor Perspectives in Medicine %R 101101/cshperspecta007039 2

  43. Palker TJ, Matthews TJ, Langlois A, Tanner ME, Martin ME, Scearce RM, Kim JE, Berzofsky JA, Bolognesi DP, Haynes BF (1989) Polyvalent human immunodeficiency virus synthetic immunogen comprised of envelope gp120 T helper cell sites and B cell neutralization epitopes. J Immunol 142:3612–3619

    CAS  PubMed  Google Scholar 

  44. Pertwee RG, Howlett AC, Abood ME, Alexander SP, Di Marzo V, Elphick MR, Greasley PJ, Hansen HS, Kunos G, Mackie K, Mechoulam R, Ross RA (2010) International union of basic and clinical pharmacology. LXXIX. Cannabinoid receptors and their ligands: beyond CB(1) and CB(2). Pharmacol Rev 62:588–631

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  45. Prentiss D, Power R, Balmas G, Tzuang G, Israelski DM (2004) Patterns of marijuana use among patients with HIV/AIDS followed in a public health care setting. J Acquir Immune Defic Syndr 35:38–45

    PubMed  Article  Google Scholar 

  46. Pross SH, Nakano Y, McHugh S, Widen R, Klein TW, Friedman H (1992) Contrasting effects of THC on adult murine lymph node and spleen cell populations stimulated with mitogen or anti-CD3 antibody. Immunopharmacol Immunotoxicol 14:675–687

    CAS  PubMed  Article  Google Scholar 

  47. Rao GK, Kaminski NE (2006) Cannabinoid-mediated elevation of intracellular calcium: a structure-activity relationship. J Pharmacol Exp Ther 317:820–829

    CAS  PubMed  Article  Google Scholar 

  48. Robinson HL, Amara RR (2005) T cell vaccines for microbial infections. Nat Med 11:S25–S32

    CAS  PubMed  Article  Google Scholar 

  49. Rosenbaum T, Simon S (2007) TRPV1 receptors and signal transduction. In: Liedtke WB, Heller S (eds) TRP Ion channel function in sensory transduction and cellular signaling cascades. CRC Press, Boca Raton

    Google Scholar 

  50. Schatz AR, Lee M, Condie RB, Pulaski JT, Kaminski NE (1997) Cannabinoid receptors CB1 and CB2: a characterization of expression and adenylate cyclase modulation within the immune system. Toxicol Appl Pharmacol 142:278–287

    CAS  PubMed  Article  Google Scholar 

  51. Springs AE, Karmaus PW, Crawford RB, Kaplan BL, Kaminski NE (2008) Effects of targeted deletion of cannabinoid receptors CB1 and CB2 on immune competence and sensitivity to immune modulation by Delta9-tetrahydrocannabinol. J Leukoc Biol 84:1574–1584

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  52. Surman S, Lockey TD, Slobod KS, Jones B, Riberdy JM, White SW, Doherty PC, Hurwitz JL (2001) Localization of CD4+ T cell epitope hotspots to exposed strands of HIV envelope glycoprotein suggests structural influences on antigen processing. Proc Natl Acad Sci U S A 98:4587–4592

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  53. Wacnik PW, Luhr KM, Hill RH, Ljunggren HG, Kristensson K, Svensson M (2008) Cannabinoids affect dendritic cell (DC) potassium channel function and modulate DC T cell stimulatory capacity. J Immunol 181:3057–3066

    CAS  PubMed  Article  Google Scholar 

  54. Walker BD, Burton DR (2008) Toward an AIDS vaccine. Science 320:760–764

    CAS  PubMed  Article  Google Scholar 

  55. Wang WK, Dudek T, Essex M, Lee TH (1999) Hypervariable region 3 residues of HIV type 1 gp120 involved in CCR5 coreceptor utilization: therapeutic and prophylactic implications. Proc Natl Acad Sci U S A 96:4558–4562

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  56. Winsauer PJ, Molina PE, Amedee AM, Filipeanu CM, McGoey RR, Troxclair DA, Walker EM, Birke LL, Stouwe CV, Howard JM, Leonard ST, Moerschbaecher JM, Lewis PB (2011) Tolerance to chronic delta-9-tetrahydrocannabinol (Delta-THC) in rhesus macaques infected with simian immunodeficiency virus. Exp Clin Psychopharmacol 19:154–172

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  57. Zhu LX, Sharma S, Stolina M, Gardner B, Roth MD, Tashkin DP, Dubinett SM (2000) Delta-9-tetrahydrocannabinol inhibits antitumor immunity by a CB2 receptor-mediated, cytokine-dependent pathway. J Immunol 165:373–380

    CAS  PubMed  Article  Google Scholar 

  58. Zygmunt PM, Petersson J, Andersson DA, Chuang H, Sorgard M, Di Marzo V, Julius D, Hogestatt ED (1999) Vanilloid receptors on sensory nerves mediate the vasodilator action of anandamide. Nature 400:452–457

    CAS  PubMed  Article  Google Scholar 

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Acknowledgments

The authors thank Jose Suarez and Natalia Kovalova for technical support during the study, and Kimberly Hambleton for assistance with submission of the manuscript.

Funding

This study was funded by National Institutes of Health (grant number DA007908).

Conflict of Interests

The authors declare that they have no conflict of interest.

Authorship Contributions

• W Chen, BLF Kaplan, and NE Kaminski participated in research design

• W Chen and RB Crawford conducted experiments

• W Chen performed data analysis

• W Chen wrote the paper

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Correspondence to Norbert E. Kaminski.

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Chen, W., Crawford, R.B., Kaplan, B.L.F. et al. Modulation of HIVGP120 Antigen-Specific Immune Responses In Vivo by Δ9-Tetrahydrocannabinol. J Neuroimmune Pharmacol 10, 344–355 (2015). https://doi.org/10.1007/s11481-015-9597-x

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Keywords

  • Cannabinoids
  • Cannabinoid receptors
  • HIVgp120
  • Immune modulation