Correspondence

Kaposi's Sarcoma and Herpesvirus-Like DNA Sequences in Sensory Ganglia

N Engl J Med 1996; 334:1341May 16, 1996

Article

To the Editor:

Human herpesvirus 8 (HHV-8), also known as Kaposi's sarcoma–associated herpesvirus (KSHV), has recently been proposed as a possible cofactor in the development of Kaposi's sarcoma.1 Specific sequences of the virus were detected by the polymerase chain reaction (PCR) in over 90 percent of Kaposi's sarcoma lesions, regardless of their association with the human immunodeficiency virus (HIV).2 Nevertheless, except for the documented tropism of KSHV for CD19-positive circulating B lymphocytes,3 little is known about sites of viral latency and persistence in infected hosts.

In immunocompromised patients, Kaposi's sarcoma frequently presents as disseminated lesions with a varicelliform distribution (i.e., a predilection for areas of the face and limbs), whereas in immunocompetent subjects the acral skin surfaces (i.e., the upper and lower extremities) are most frequently involved. Moreover, Kaposi's sarcoma with a dermatome-like distribution has been reported at the site of previous cutaneous herpes zoster.4 This prompted us to explore whether KSHV, like the alphaherpesviruses, could be detected in the peripheral nervous systems of patients with Kaposi's sarcoma and AIDS.

Seven paravertebral sensory lumbar ganglia were collected at autopsy from seven patients with AIDS who had Kaposi's sarcoma at the time of their deaths or earlier. Three specimens were from subjects described in another study.5 As negative controls we used seven paravertebral sensory ganglia obtained at autopsy from four patients with AIDS and three persons who were seronegative for HIV type 1 and had no history of Kaposi's sarcoma.

After their collection, the specimens were embedded under sterile conditions and immediately snap-frozen in liquid nitrogen. Part of each sample was processed for microscopical and immunohistochemical analysis, and part underwent digestion with proteinase K, followed by DNA extraction by a phenol–chloroform method. Microscopical and immunohistochemical analyses for CD34–positive cells were performed in each specimen to rule out the presence of occult Kaposi's sarcoma lesions. Pathological and molecular analyses were conducted in separate laboratories. In addition, PCR studies were performed in a blinded fashion.

Five hundred nanograms of DNA, as determined spectrophotometrically, were loaded into each amplification reaction. The samples were tested by a 35-cycle, hot-start PCR, followed by Southern blot hybridization of the amplified products or nested PCR (35 plus 35 cycles). Rigid measures were taken to prevent the carryover of DNA during PCR. In particular, throughout the procedure, after the study of each sample obtained at autopsy a sample of double-distilled water was studied as a negative control. DNA integrity was determined by amplifying a human β-globin gene fragment (data not shown). Details of the processing of specimens, pathological studies, and PCR are reported elsewhere.5

KSHV DNA sequences were detected by both simple and nested PCR in the paravertebral ganglia of all seven patients with AIDS and Kaposi's sarcoma (Figure 1Figure 1Nested PCR Amplification of HHV-8 DNA from Paravertebral Sensory Ganglia Obtained at Autopsy.). No specific pathological finding was associated with positivity on PCR, since the ganglia were histologically normal (Figure 2Figure 2Paravertebral Ganglion and Satellite Cells Obtained at Autopsy from a Patient with AIDS and Kaposi's Sarcoma (Hematoxylin and Eosin, ×250).). In addition, there was no evident relation between the presence of viral sequences and the stage of Kaposi's sarcoma at death. By contrast, none of the ganglia from any of the control patients were found to contain the virus.

These results extend our previous postmortem findings in patients with AIDS and Kaposi's sarcoma, in whom extralesional detection of HHV-8 was confined to secondary lymphoid organs (i.e., the spleen and the lymph nodes), the prostate gland, bone marrow, and Kaposi's sarcoma–free skin.5 Moreover, the high rate of KSHV detection in paravertebral ganglia argues for the concept of a selective tropism of the virus in the nervous system and could explain the clinical distribution of Kaposi's sarcoma cutaneous lesions. It is noteworthy that several areas of the brain cortex, basal ganglia, brain stem, and spinal cord in these patients were uniformly negative for KSHV by both simple and nested PCR.5

Issues that are still being studied include the extent of HHV-8 colonization of the sensitive and autonomic ganglia in patients with Kaposi's sarcoma, the topographic relation of such colonization to Kaposi's sarcoma lesions, the frequency of this finding in other forms of the disease, and the identity of the cellular elements that harbor the virus in the ganglia.

Mario Corbellino, M.D.
University of Milan

Carlo Parravicini, M.D.
Ospedale Luigi Sacco, 20157 Milan, Italy

Jean Thierry Aubin, Ph.D.
Hôpital Pitié–Salpêtrière, 75013 Paris, France

Emilio Berti, M.D.
Ospedale Maggiore–Policlinico, 20100 Milan, Italy

5 References

1. 1

   Chang Y, Cesarman E, Pessin MS, et al. Identification of herpesvirus-like DNA sequences in AIDS-associated Kaposi's sarcoma. Science 1994;266:1865-1869
   CrossRef | Web of Science | Medline

2. 2

   Moore PS, Chang Y. Detection of herpesvirus-like DNA sequences in Kaposi's sarcoma in patients with and those without HIV infection. N Engl J Med 1995;332:1181-1185
   Full Text | Web of Science | Medline

3. 3

   Ambroziak JA, Blackbourn DJ, Herndier BG, et al. Herpes-like sequences in HIV-infected and uninfected Kaposi's sarcoma patients. Science 1995;268:582-583
   CrossRef | Web of Science | Medline

4. 4

   Niedt GW, Prioleau PG. Kaposi's sarcoma occurring in a dermatome previously involved by herpes zoster. J Am Acad Dermatol 1988;18:448-451
   CrossRef | Web of Science | Medline

5. 5

   Corbellino M, Poirel L, Bestetti G, et al. Restricted tissue distribution of extra-lesional KS-associated herpesvirus-like (KSHV) DNA sequences in AIDS patients with Kaposi's sarcoma. AIDS Res Hum Retroviruses (in press).
   

Citing Articles (12)

Citing Articles

1. 1

   Upton D. Allen. (2002) Human herpesvirus type 8 infections among solid organ transplant recipients. Pediatric Transplantation 6:3, 187-192
   CrossRef

2. 2

   L. Brambilla, V. Boneschi, S. Ferrucci, M. Taglioni, E. Berti. (2000) Human herpesvirus-8 infection among heterosexual partners of patients with classical Kaposi’s sarcoma. British Journal of Dermatology 143:5, 1021-1025
   CrossRef

3. 3

   Paul K.S. Chan, Ho-Keung Ng, Mamie Hui, Margaret Ip, Jo L.K. Cheung, Augustine F. Cheng. (1999) Presence of human herpesviruses 6, 7, and 8 DNA sequences in normal brain tissue. Journal of Medical Virology 59:4, 491-495
   CrossRef

4. 4

   Angelo V. Marzano, Emilio Berti, Elvio Alessi. (1999) Primary cutaneous B-cell lymphoma with a dermatomal distribution. Journal of the American Academy of Dermatology 41:5, 884-886
   CrossRef

5. 5

   Silvia Fossati, Vinicio Boneschi, Silvia Ferrucci, Lucia Brambilla. (1999) Human immunodeficiency virus negative Kaposi sarcoma and lymphoproliferative disorders. Cancer 85:7, 1611-1615
   CrossRef

6. 6

   Pfrommer, Tebbe, Tidona, WOlfer, Krengel, Zeichardt, Zouboulis, Orfanos. (1998) Progressive HHV-8-positive classic Kaposi's sarcoma: rapid response to interferon alpha-2a but persistence of HHV-8 DNA sequences in lesional skin. British Journal of Dermatology 139:3, 516-519
   CrossRef

7. 7

   Kamal G. Ishak, Karen E. Bijwaard, Hala R. Makhlouf, Jeffery K. Taubenberger, Jack H. Lichy, Zachary D. Goodman. (1998) Absence of human herpesvirus 8 DNA sequences in vascular tumors of the liver*. Liver 18:2, 124-127
   CrossRef

8. 8

   Ruth M. Greenblatt. (1998) KAPOSI'S SARCOMA AND HUMAN HERPESVIRUS-8. Infectious Disease Clinics of North America 12:1, 63-82
   CrossRef

9. 9

   M DICTOR. (1997) Human herpesvirus 8 and Kaposi's sarcoma1. Seminars in Cutaneous Medicine and Surgery 16:3, 181-187
   CrossRef

10. 10

    E. BERTI, A.V MARZANO, I. DECLEVA, G. BESTETTI, M. PIZZUTO, L. POIREL, M. CAPRA, C. PARRAVICINI, M. CORBELLINO. (1997) Simultaneous onset of primary cutaneous B-cell lymphoma and human herpesvirus 8-associated Kaposi's sarcoma. British Journal of Dermatology 136:6, 924-929
    CrossRef

11. 11

    Jay A Levy. (1997) Three new human herpesviruses (HHV6, 7, and 8). The Lancet 349:9051, 558-563
    CrossRef

12. 12

    (1996) Absence of HHV-8 in Prostate and Semen. New England Journal of Medicine 335:16, 1237-1239
    Full Text