1887

Chapter 95 : General Immunologic Evaluation of Patients with Human Immunodeficiency Virus Infection

MyBook is a cheap paperback edition of the original book and will be sold at uniform, low price.

Preview this chapter:
Zoom in
Zoomout

General Immunologic Evaluation of Patients with Human Immunodeficiency Virus Infection, Page 1 of 2

| /docserver/preview/fulltext/10.1128/9781555815905/9781555813642_Chap95-1.gif /docserver/preview/fulltext/10.1128/9781555815905/9781555813642_Chap95-2.gif

Abstract:

This chapter provides proven methods for the routine immunologic monitoring of the cellular immune status of patients with human immunodeficiency virus (HIV) infection. Since a majority of immunologic assays are performed with peripheral blood mononuclear cells (PBMCs), procedures for the isolation and cryopreservation of PBMCs are also discussed in the chapter. The section describing each procedure is prefaced with an explanation of the rationale behind the described assay, quality control measures necessary for standardization of testing, and practical advice on pitfalls to avoid. When monitoring the immunologic status of patients with HIV infection, isolation of lymphoid cells is often required for performing several immunologic procedures, such as lymphocyte immunophenotyping and measurement of lymphocyte function. Three- and four-color panels of monoclonal antibodies that may be used for the basic monitoring of patients with AIDS have also been discussed. In summary, immunophenotyping of patients with HIV infection can provide the most clinically relevant information as to the immunologic status of these patients.

Citation: Stevens R, Lempicki R, Natarajan V, Higgins J, Adelsberger J, Metcalf J. 2006. General Immunologic Evaluation of Patients with Human Immunodeficiency Virus Infection, p 847-861. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch95
Highlighted Text: Show | Hide
Loading full text...

Full text loading...

Figures

Image of FIGURE 1
FIGURE 1

Ex vivo methods of cell turnover are well correlated. (A) Ex vivo BrdU incorporation levels for CD4 and CD8 T cells correlated in paired samples from freshly isolated blood and lymph node suspensions from 14 subjects infected with HIV-1; (B) ex vivo BrdU incorporation correlated with Ki67 staining using the same blood samples from 56 individuals that were either healthy controls or infected with HIV; (C) ex vivo BrdU incorporation correlated with Ki67 staining in CD4 T cells from longitudinal blood samples taken once every day or two from three HIV-infected subjects.

Citation: Stevens R, Lempicki R, Natarajan V, Higgins J, Adelsberger J, Metcalf J. 2006. General Immunologic Evaluation of Patients with Human Immunodeficiency Virus Infection, p 847-861. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch95
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of FIGURE 2
FIGURE 2

Ex vivo BrdU incorporation and Ki67 staining methods correlated with in vivo BrdU labeling. (A) Ex vivo BrdU incorporation levels in CD4 and CD8 T cells determined for 14 HIV-infected subjects just before a 30-min in vivo BrdU administration were shown to correlate with peak in vivo BrdU incorporation levels; (B) Ki67 T-cell staining levels were found to correlate with the in vivo BrdU T-cell incorporation levels in six HIV-infected subjects infused with BrdU.

Citation: Stevens R, Lempicki R, Natarajan V, Higgins J, Adelsberger J, Metcalf J. 2006. General Immunologic Evaluation of Patients with Human Immunodeficiency Virus Infection, p 847-861. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch95
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of FIGURE 3
FIGURE 3

In vivo BrdU incorporation correlates with in vivo deuterated-glucose labeling in a double-labeling study. A 30-min infusion of BrdU and a 5-day continuous infusion of deuterated glucose were administered to an HIV-infected subject. (A) Samples were assayed for label incorporation and decay in CD4 T cells over a 1.5-year period; (B) in vivo BrdU incorporation and decay were highly correlated with incorporation and decay levels of deuterated glucose.

Citation: Stevens R, Lempicki R, Natarajan V, Higgins J, Adelsberger J, Metcalf J. 2006. General Immunologic Evaluation of Patients with Human Immunodeficiency Virus Infection, p 847-861. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch95
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of FIGURE 4
FIGURE 4

Quantitation of TRECs in patient samples. Shown is a standard curve plotting log starting TREC copy number versus threshold cycle. The amounts of TREC in unknown samples were calculated from the standard curve by using ABI Prism 7700 software.

Citation: Stevens R, Lempicki R, Natarajan V, Higgins J, Adelsberger J, Metcalf J. 2006. General Immunologic Evaluation of Patients with Human Immunodeficiency Virus Infection, p 847-861. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch95
Permissions and Reprints Request Permissions
Download as Powerpoint

References

/content/book/10.1128/9781555815905.ch95
1. Bruno, S., and , Z. Darzynkiewicz. 1992. Cell cycle dependent expression and stability of the nuclear protein detected by Ki-67 antibody in HL-60 cells. Cell Prolif. 25:3140.
2. Centers for Disease Control and Prevention. 1997. 1997 revised guidelines for the performance of CD4+ T-cell determination in persons infected with human immunodeficiency virus (HIV). Morb. Mortal. Wkly. Rep. 46(RR-2): 129.
3. Centers for Disease Control and Prevention. 2002. Guidelines for preventing opportunistic infections among HIVinfected persons—2002. Morb. Mortal. Wkly. Rep. 51 (RR-8):146.
4. Centers for Disease Control and Prevention. 2002. Guidelines for using antiretroviral agents among HIVinfected adults and adolescents. Morb. Mortal. Wkly. Rep. 51(RR-7):171.
5. Centers for Disease Control and Prevention. 2003. 2003 guidelines for performing single-platform absolute CD4+ T-cell determinations with CD45 gating for persons infected with human immunodeficiency virus (HIV). Morb. Mortal. Wkly. Rep. 52(RR-2):113.
6. Douek, D. C.,, R. D. McFarland,, P. H. Keiser,, E. A. Gage,, J. M. Massey,, B. F. Haynes,, M. A. Polis,, A. T. Haase,, M. B. Feinberg,, J. L. Sullivan,, B. D. Jamieson,, J. A. Zack,, L. J. Picker, and , R. A. Koup. 1998. Changes in thymic function with age and during the treatment of HIV infection. Nature 396:690695.
7. Effros, R. B.,, R. Allsopp,, C. P. Chiu,, M. A. Hausner,, K. Hirji,, L. Wang,, C. B. Harley,, B. Villeponteau,, M. D. West, and , J. V. Giorgi. 1996. Shortened telomeres in the expanded CD28— CD8+ cell subset in HIV disease implicate replicative senescence in HIV pathogenesis. AIDS 10:1722.
8. Gerdes, J.,, H. Lemke,, H. Baisch,, H. H. Wacker,, U. Schwab, and , H. Stein. 1984. Cell cycle analysis of a cell proliferation-associated human nuclear antigen defined by the monoclonal antibody Ki-67. J. Immunol. 133:17101715.
9. Heid, C. A.,, J. Stevens,, K. J. Livak, and , P. M. Williams. 1996. Real time quantitative PCR. Genome Res. 6:986994.
10. Hellerstein, M.,, M. B. Hanley,, D. Cesar,, S. Siler,, C. Papageorgopoulos,, E. Wieder,, D. Schmidt,, R. Hoh,, R. Neese,, D. Macallan,, S. Deeks, and , J. M. McCune. 1999. Directly measured kinetics of circulating T lymphocytes in normal and HIV 1-infected humans. Nat. Med. 5:8389.
11. Hellerstein, M. K.,, R. A. Hoh,, M. B. Hanley,, D. Cesar,, D. Lee,, R. A. Neese, and , J. M. McCune. 2003. Subpopulations of long-lived and short-lived T cells in advanced HIV-1 infection. J. Clin. Investig. 112:956966.
12. Kausch, I.,, A. Lingnau,, E. Endl,, K. Sellmann,, I. Deinert,, T. L. Ratliff,, D. Jocham,, G. Sczakiel,, J. Gerdes, and , A. Bohle. 2003. Antisense treatment against Ki-67 mRNA inhibits proliferation and tumor growth in vitro and in vivo. Int. J. Cancer 105:710716.
13. Koenig, S., and , A. S. Fauci. 1988. AIDS: immunopatho-genesis and immune response to the human immunodeficiency virus, p. 6177. In V T. DeVita, Jr.,, S. Hellman, and , S. A. Rosenberg (ed.), AIDS: Etiology, Diagnosis, Treatment, and Prevention, 2nd ed. J. B. Lippincott Co., Philadelphia, Pa.
14. Koopman, G.,, C. P. M. Reutelingsperger,, G. A. M. Kuijten,, R. M. J. Keehnen,, S. T. Pals, and , M. H. J. van Oers. 1994. Annexin V for flow cytometric detection of phos-phatidylserine expression of B cells undergoing apoptosis. Blood 84:14151420.
15. Kovacs, J. A.,, R. A. Lempicki,, I. A. Sidorov,, J. W. Adelsberger,, B. Herpin,, J. A. Metcalf,, I. Sereti,, M. A. Polis,, R. T. Davey,, J. Tavel,, J. Falloon,, R. Stevens,, L. Lambert,, R. Dewar,, D. J. Schwartzentruber,, M. R. Anver,, M. W. Baseler,, H. Masur,, D. S. Dimitrov, and , H. C. Lane. 2001. Identification of dynamically distinct subpopulations of T lymphocytes that are differentially affected by HIV. J. Exp. Med. 194:17311741.
16. Lane, H. C.,, J. M. Depper,, W. C. Green,, G. Whalen,, T. A. Waldmann, and , A. S. Fauci. 1985. Qualitative analysis of immune function in patients with acquired immunodeficiency syndrome: evidence for a selective defect in soluble antigen recognition. N Engl. J. Med. 313:7984.
17. Lane, H. C., and , A. S. Fauci. 1985. Immunologic abnormalities in the acquired immunodeficiency syndrome. Annu. Rev. Immunol. 3:477500.
18. Lempicki, R. A.,, J. A. Kovacs,, M. W. Baseler,, J. W. Adelsberger,, R. L. Dewar., V. Natarajan,, M. C. Bosche,, J. A. Metcalf,, R. A. Stevens,, L. A. Lambert,, W. G. Alvord,, M. A. Polis,, R. T. Davey,, D. S. Dimitrov, and , H. C. Lane. 2000. Impact of HIV-1 infection and highly active antiretroviral therapy on the kinetics of CD4+ and CD8+ T cell turnover in HIVinfected patients. Proc. Natl. Acad. Sci. USA 97:1377813783.
19. McFarland, R. D.,, D. C. Douek,, R. A. Koup, and , L. J. Picker. 2000. Identification of a human recent thymic emigrant phenotype. Proc. Natl. Acad. Sci. USA 97:42154220.
20. Natarajan, V.,, R. A. Lempicki,, I. Sereti,, Y. Badralmaa,, J. W. Adelsberger,, J. A. Metcalf,, D. A. Prieto,, R. Stevens,, M. W. Baseler,, J. A. Kovacs, and , H. C. Lane. 2002. Increased peripheral expansion of naive CD4+T cells in vivo after IL-2 treatment of patients with HIV infection. Proc. Natl. Acad. Sci. USA 99:1071210717.
21. National Committee for Clinical Laboratory Standards. 1989. Clinical Applications for Flow Cytometry: Quality Assurance and Immunophenotyping of Peripheral Blood Lymphocytes; Proposed Guideline. NCCLS document H42-P. National Committee for Clinical Laboratory Standards, Villanova, Pa.
22. Reimann, K. A.,, M. R. G. O’Gorman,, J. Spritzler,, C. L. Wilkening,, D. E. Sabath,, K. Helm,, D. E. Campbell, and The NIAID DAIDS New Technologies Evaluation Group. 2000. A multisite comparison of CD4 and CD8 T lymphocyte counting by single vs. multiple platform methodologies: evaluation of Beckman Coulter Flow-Count Fluorospheres and tetraONE System. Clin. Diagn. Lab. Immunol. 7:344351.
23. Sachsenberg, N.,, A. S. Perelson,, S. Yerly,, G. A. Schockmel,, D. Leduc,, B. Hirschel, and , L. Perrin. 1998. Turnover of CD4(+) and CD8(+) T lymphocytes in HIV-1 infection as measured by Ki-67 antigen. J. Exp. Med. 187:12951303.
24. Schnizlein-Bick, C. T.,, J. Spritzler,, C. L. Wilkening,, J. K. A. Nicholson,, M. R. G. O’Gorman,Site Investigators, and The NIAID DAIDS New Technologies Evaluation Group. 2000. Evaluation of TruCount absolute-count tubes for determining CD4 and CD8 cell numbers in human immunodeficiency virus-positive adults. Clin. Diagn. Lab. Immunol. 7:336343.
25. Strong, D. M.,, J. R. Ortaldo,, F. Pandolfi,, A. Maluish, and , R. B. Herberman. 1982. Cryopreservation of human mononuclear cells for quality control in clinical immunology. I. Correlations in recovery of K- and NK-cell functions, surface markers and morphology. J. Clin. Immunol. 2:216221.
26. UNAIDS. 2004. 2004 Report on the Global AIDS Epidemic: 4th Global Report. UNAIDS, Geneva, Switzerland.

Tables

Generic image for table
TABLE 1

Suggested monoclonal antibody panels for lymphocyte immunophenotyping of patients with HIV infection

Citation: Stevens R, Lempicki R, Natarajan V, Higgins J, Adelsberger J, Metcalf J. 2006. General Immunologic Evaluation of Patients with Human Immunodeficiency Virus Infection, p 847-861. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch95
Generic image for table

Citation: Stevens R, Lempicki R, Natarajan V, Higgins J, Adelsberger J, Metcalf J. 2006. General Immunologic Evaluation of Patients with Human Immunodeficiency Virus Infection, p 847-861. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch95

This is a required field
Please enter a valid email address
Please check the format of the address you have entered.
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error