Oral and systemic factors associated with increased levels of human immunodeficiency virus type 1 RNA in saliva

doi:10.1016/S1079-2104(00)70124-7

Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology

Volume 89, Issue 4, April 2000, Pages 432-440

https://doi.org/10.1016/S1079-2104(00)70124-7 Get rights and content

Abstract

Objective. The purpose of this investigation was to quantify human immunodeficiency virus type-1 (HIV-1) RNA in saliva and plasma and identify factors associated with increased salivary viral load. Study design. Forty HIV-1–seropositive adults underwent oral examinations to assess mucosal and periodontal health. Whole saliva was evaluated for HIV-1 RNA titer and occult blood. Plasma viral load, CD4 cell count, HIV-1 staging, and antiretroviral therapy data were obtained from medical records. Associations between salivary titers and oral/systemic parameters were analyzed by means of t tests, Wilcoxon signed rank tests, Pearson’s correlation coefficient, and analysis of covariance. Results. Forty-two percent of the subjects had detectable salivary HIV-1 RNA. Oral titers were highly correlated with plasma viral levels (r = 0.51, P < .01). HIV-associated periodontal disease (in particular, linear gingival erythema), severe gingival inflammation, and absence of antiretroviral therapy were associated with high salivary titers (P < .01). Conclusions. Substantial quantities of HIV-1 can be shed in the oral cavity, particularly when inflammatory conditions are present. Salivary titer may be a useful indicator of systemic viral burden. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2000;89:432-40)

Section snippets

Subjects

Forty HIV-1 seropositive adults were recruited from an ongoing investigation of HIV-1 disease and oral health in the southeastern United States. Subjects were referred from the Infectious Diseases Clinic at the University of North Carolina (UNC) Health Care system. Study eligibility included racial self-identification as either Caucasian (non-Hispanic) or African-American, at least 3 posterior teeth in each quadrant of the mouth, and a plasma viral load ascertained within 8 weeks of study visit

HIV-1 spiking experiment

To verify the suitability of the NASBA technique for viral RNA quantitation of oral fluids, saliva and plasma from an uninfected donor were spiked in triplicate with the HIV-1 Ba-L isolate at one of two concentrations and then analyzed for viral load through use of the Nuclisens assay. As shown in Fig 1, similar results were obtained when virus was spiked into saliva or plasma at the concentrations tested (P > .05, t test), confirming the lack of saliva-mediated inhibition in the Nuclisens

Discussion

This cross-sectional investigation examined associations between elevated HIV-1 RNA levels in saliva and viral, immunologic, and oral health parameters. These parameters included 4 measures of systemic disease (plasma viral load, CD4 cell count, AIDS diagnosis, and ART) and 6 measures of oral health (HIV-associated mucosal lesions, occult salivary blood, HIV-related periodontal diseases, pocket probing depth, clinical attachment level, and gingival inflammation). We demonstrate that the saliva

Acknowledgements

We thank Dawn Rogers for patient recruitment and saliva collection, Jody Shock for viral load assays, and Audrey Alexander and Stephanie Freel for technical assistance.

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The processes and interactions that occur following co-infection with viral and bacterial pathogens are complex, and can lead to a protracted disease course. P. gingivalis infection can induce the reactivation of Epstein-Barr (EB) virus and HIV (Contreras, Botero, & Slots, 2014; Verdugo et al., 2012), which is potentially important because Epstein-Barr virus and HIV pre-viral DNA and RNA has a complex relationship with severe periodontal disease (Shugars, Slade, Patton, & Fiscus, 2000; Slots, 2010). It has been found that P. gingivalis can induce latent infection in the host cell by HIV-1 activation as a result of chromatin modifications (Imai, Ochiai, & Okamoto, 2009; Niederman, Buyle-Bodin, Lu, Robinson, & Naleway, 1997).
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An MTT assay revealed that cell viability was significantly lower in the IAV group than in the P. gingivalis + IAV group (P < 0.05). Flow cytometry indicated that the apoptosis rate was significantly higher in the IAV group than in the P. gingivalis + IAV group (P < 0.05). The fluorescence levels of GFP-LC3 increased significantly, the LC3-II/LC3-I ratio was significantly higher, and the P62 protein levels were statistically lower in the P. gingivalis + IAV group compared with the IAV group (all P < 0.05). Western blotting revealed that the LC3- II/LC3-I ratio was significantly lower, and caspase-3 levels were significantly higher in the 3MA + P. gingivalis + IAV group compared to the P. gingivalis + IAV group (all P < 0.05).
In vitro studies showed that infection by P. gingivalis combined with IAV temporarily inhibited apoptosis in respiratory epithelial cells, and this may be related to the initiation of autophagy.
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This is particularly true in HIV patients who have severe periodontal disease, in which the milieu of pathogenic bacterial infection and chronic inflammation appears to enhance HIV replication. Not only are more periodontal pathogens such as Pg found in HIV-positive individuals than in healthy controls (Chattin et al., 1999; Scully et al., 1999), but there was a significant correlation between the clinical stage of periodontitis and HIV-1 proviral DNA load in the gingival crevicular fluid and HIV-1 viral RNA load in the plasma and saliva (Maticic et al., 2000; Shugars et al., 2000). The interactions between periodontal pathogens and HIV are therefore likely to create a vicious cycle of enhanced viral induction and bacterial replication in the oral cavity of HIV patients.
HIV patients with severe periodontitis have high levels of residual virus in their saliva and plasma despite effective therapy (HAART). Multiple short chain fatty acids (SCFAs) from periodontal pathogens reactivate HIV-1 in both Jurkat and primary T-cell models of latency. SCFAs not only activate positive transcription elongation factor b (P-TEFb), which is an essential cellular cofactor for Tat, but can also reverse chromatin blocks by inducing histone modifications. SCFAs simultaneously increase histone acetylation by inhibiting class-1/2 histone deacetylases (HDACs) and decrease repressive histone tri-methylation at the proviral LTR by downregulating expression of the class-3 HDAC sirtuin-1 (SIRT1), and the histone methyltransferases enhancer of Zeste homolog 2 (EZH2) and suppressor of variegation 3–9 homolog 1 (SUV39H1). Our findings provide a mechanistic link between periodontal disease and enhanced HIV-1 replication, and suggest that treatment of periodontal disease, or blocking the activities of SCFAs, will have a therapeutic benefit for HIV patients.
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^☆: Reprint requests: Diane C. Shugars, DDS, MPH, PhD, Department of Dental Ecology, UNC School of Dentistry CB#7450, University of North Carolina, Chapel Hill, NC 27599-7450

^☆☆: This study was supported by the National Institutes of Health (R01DE12162, R29DE11369) and the UNC Center for AIDS Research (NIH P30HD37260).

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Oral MedicineOral and systemic factors associated with increased levels of human immunodeficiency virus type 1 RNA in saliva☆,☆☆

Abstract

Section snippets

Subjects

HIV-1 spiking experiment

Discussion

Acknowledgements

J Am Dent Assoc

J Virol Methods

J Am Dent Assoc

Oral Surg Oral Med Oral Pathol

Lancet

HIV viral load markers in clinical practice

Nature Med

Viral counts count in HIV infection

Science

Maternal plasma human immunodeficiency virus type 1 RNA level: a determinant and projected threshold for mother-to-child transmission

Proc Natl Acad Sci U S A

Biological correlates of HIV-1 heterosexual transmission

AIDS

Prognosis in HIV-1 infection predicted by the quantity of virus in plasma

Science

Analysis of HIV-1 load in plasma, semen and saliva: evidence for different viral compartments in a cross-sectional and longitudinal study

AIDS

Quantification of HIV in semen: correlation with antiviral treatment and immune status

AIDS

Association between culturable human immunodeficiency virus type 1 (HIV-1) in semen and HIV-1 RNA levels in semen and blood: evidence for compartmentalization of HIV-1 between semen and blood

J Inf Dis

Cellular and anatomical reservoirs of HIV-1 in patients receiving potent antiretroviral combination therapy

JAMA

Human immunodeficiency virus type 1 and cytomegalovirus in saliva

J Med Virol

Low level of cell-free virus detected at high frequency in saliva from HIV-1-infected individuals

AIDS

Oral HIV-1 recovery in the presence of periodontal disease

Oral Dis

The anti-HIV-1 activity associated with saliva

J Dent Res

Patients with human immunodeficiency virus type 1 have low levels of virus in saliva even in the presence of periodontal disease

J Infect Dis

Quantitation of HIV-1 RNA from different biological compartments.

The one-tube quantitative HIV-1 RNA NASBA: precision, accuracy and application

PCR Methods Appl

Quantification of HIV-1 RNA in plasma: comparable results with the NASBA HIV-1 RNA QT and the AMPLICOR HIV Monitor Test

J Acquir Immune Defic Syndr Hum Retrovirol

Oral Medicine
Oral and systemic factors associated with increased levels of human immunodeficiency virus type 1 RNA in saliva☆,☆☆