Bridging the Gap: Conference Record [Abstract book, International Conference on AIDS (12th: 1998: Geneva, Switzerland)]

540 Abstracts 31223-32101

12th World AIDS Conference

alone and 1 each by E.coli and Salmonella enteritidis. Amongst 25 immunocompromised patients 6 had Cryptosporidium, 1 had Ascaris, 1 had Ascaris with
Trichuris and 1 had Strongyloidis. Among 25 healthy controls, 2 had Ascaris and
1 had Trichuris and 1 had Hookworm.
Conclusion: Though AIDS patients are easily affected by all opportunistic
infections, the most common cause of diarrhoea in Calcutta, India seems to be
Cryptosporidium.
31223 Modulation of DNA viruses regulatory regions by HIV-1
Tat: Transactivation mechanisms of HPV-16 long control
region (LCR)
Marialina Tornesello, F.M. Buonaguro1, A. Meglio1, L. Buonaguro',
E. Beth-Giraldo', G. Giraldo1. Viral Oncology - 1Istituto Nazionale Tumori "Fond.
Pascale", Cappellacangiani, Naples, Italy
Background: Since the HIV/AIDS epidemic three neoplasia (Kaposi's sarcoma,
lymphomas, and ano-genital dysplasia) previously associated to Herpesviruses
and Papillomaviruses, respectively, have shown a relevant incidence increase.
Clnical/epidemiological observations suggest that HIV or the HIV-related immunosuppression plays a role in the etiopathogenesis of such lesions. The higher
incidence of HPV infection and HPV-associated lesions at all levels of immune
suppression in HIV-positive subjects, however, provide a stronger evidence for
the HIV direct role on the natural history of viral infections. We are analyzing a)
the possible role of HIV on HPV gene expression and/or virus reactivation and
b) the molecular mechanisms involved in the interaction of HIV with heterologous
promoters of DNA viruses.
Methods and Results: We have previously shown that HIV-1 Tat protein transactivates HPV-16 regulatory regions (LCR) (Tornesello M.L. et al., Intervirology
1993) and increases the HPV-16 E6 and E7 transforming activity (Buonaguro F.M.
et al., Antibiotics and Chemother 1994). More recently we reported that HPV-16
LCR transactivation is cell-type specific and the mechanism involved seems to
be distinct from the TPA-dependent PKC signal cascade. Furthermore we have
observed that HIV-Tat is also able to transactivate the early promoter/enhancer of
the SV40 virus, another member of the Papovaviridae family. In order to analyze
the indirect mechanisms of Tat-dependent transactivation of Papovavirus regulatory regions, in particular intranuclear modulation of nuclear transcription factors,
several biomolecular approaches have been undertaken.
Papovavirus Tat responsive regions. To identify the Tat responsive elements
within the SV40 and HPV 16 regulatory regions recombinant plasmids containing deletion mutants of the HPV16 enhancer (nt 7524-7770) and HPV16 P97
promoter (nt 7778 to 1 to 57), and deletion mutants of SV40 control elements
(nt 5171 to 1 to 347), driving a cat reporter gene, have been constructed and
transiently transfected into several cell lines (HeLa, SiHa, HT3, NTERA-2), with
or without Tat stimulation. Our results show that Tat does not modulate the HPV
and the SV40 promoter regions, on the contrary Tat treatment transactivates the
HPV-16 and SV40 enhancer regions inducing a 3-8 and a 4-5 fold Cat expression
increase, respectively.
Tat-inducible nuclear factors. To identify specific Tat-modulated nuclear factors,
which in turn could influence the HPV-16 and the SV40 expression, band shift
assays have been performed with cellular extracts from Tat-treated epithelial cells.
The results obtained so far show that Tat protein is able to induce, in all tested cell
lines, the NFkB nuclear factor, whose binding sites are present within HIV LTR but
absent on the HPV LCR and SV40 regulatory regions. No modulation is observed
on AP-1 nuclear factors, whose binding sites are present within enhancer regions
of both viruses. We are currently testing the remaining nuclear factors which bind
to both HPV and SV40 regulatory regions.
Conclusions: We have been accumulating circumstantial evidence on the HIV
Tat transactivation of DNA virus regulatory regions. Data obtained so far suggest
that: a) Tat transactivates HPV-16 LCR; b) the transactivation affects also the
expression of another member of Papovavirus, the SV40; c) modulation of both
viruses is cell-type specific, being observed prevalently in epithelial cells. The
working hypothesis that we are testing is that Tat protein is able to alter the
intracellular pool of cell-specific nuclear factors which in turn activates cell and
infecting-virus gene expression.
Supported by Ministero Italiano della Sanita (Ric. Corrente 1998), X Prog.
AIDS-ISS (1998), and the ICSC-World Lab, Lausanne (Project MCD-2/7).
| 31224 0 Q-fever associated granulomatous skin lesions in a
patient with acquired immune deficiency syndrome
Yasmin Arikan1, David Burdge1, W.A. McLeod2. 1Oak Tree Clinic and the
University of British of Columbia, Oak Tree Clinic and The University of British
Columbia, Vancouver, BC; 2St. Paul's Hospital and the University of British
Columbia, Vancouver, BC, Canada
Background: Immunosuppressed patients with HIV/AIDS may have atypical
presentations of non-AIDS defining infections. There has been no previous published information regarding Q-fever in HIV/AIDS. In normal hosts, disseminated
granulomatous skin lesions have never been reported with Q-fever.

Methods: The patients medical records were reviewed. Phase II complement
fixation Q-fever serology was performed in a provincial reference laboratory. Skin
biopsy specimens were processed routinely in a university hospital pathology
laboratory, and also submitted to reference laboratories for mycobacteria, fungal
stains and culture, and PCR studies including for mycobacteria and Q-fever.
Results: Case Report: A 31-year-old woman with AIDS and CD4 count of 0.001
x 109/L(1) developed maculopapular rash, fever, headache, increased liver en
zymes and hepatosplenomegaly. The rash appeared "vasculitic" and involved the
extremities including the palms and soles. She had been on treatment for M.
avium complex (MAC) infection with rifampin, ethambutol and clarithromycin for
one month. Skin biopsy showed granulomas, but no vasculitis. Special studies
for mycobacteria, viruses and other pathogens were negative. Q-fever phase
II complement fixation serology was reactive (1:64) at time of presentation and
convalescent serology showed a four fold decrease in titre (1:16), both highly supportive of a diagnosis of Q-fever. The rash, hepatosplenomegaly, abnormal liver
enzymes and fever slowly improved after she started combination antiretroviral
therapy, and with ongoing clarithromycin therapy.
Conclusions: Q-fever should be considered in the differential diagnosis of
febrile syndromes associated with skin rash in patients with HIV/AIDS. Skin
biopsy can help define investigative approaches and lead to diagnosis; in our
patient granulomas were demonstrated on biopsy of a "vasculitic" appearing rash
that involved the palms and soles.
131225 1Discontinue primary prophylaxis regimens in selected
HIV-infected patients treated with HAART
Isabelle Ravaux1, S. Chadapavd2, A.N. Quinson2, V. Dahan2, H. Gallais2. 1La
Conception Hospital 147 Boulvard Baille 13005 Narseille; 2lnfection Disease
Unit Conception Hal Narseille, France
Objectives: Among patients treated with Highly Active AntiRetroviral Therapy
(HAART), is it possible to discontinue primary prophylaxis when CD4 cell count
is up to 300/mm3 and viral load (plasma HIV RNA) is below 500 copies/ml for at
least 3 months?
Design: Retrospective study.
Methods: From april 1996 to december 1997, among 595 patients with HAART,
we selected 40 of them with CD4 > 300 mm3 and vital load <500 copies,
for at least 3 months and we proposed them to stop primary prophylaxis for
Pneumocystis Carinii Pneumonia (PCP) and Toxoplasmosis (Toxo) by trimetothoprim/sulfamethoxazole.
Results: During 1997, in this selected group, 12 needed prophylaxis reintroduction because of viral load increase and/or CD4 cell count decrease. For the
others, during the last 6 months nor PCP, neither Toxo accured.
Conclusion: When a regulary clinical and biological follow up is effected, we
suggest that discontinuation of primary prophylaxis for PCP and Toxoplasmosis
is reasonable, only if HAART controle the immune deficit.
[31226 Prevalence and consequences of cryptosporidium
oocysts in drinking water
Kenndy Kaonga1, B. Kangende2. 1University Teaching Hospital, Pathology and
Microbiology, P/B RW 1X, Lusaka; 2Lusaka Water and Sewerage Company,
Zambia
Objectives: To assess the extent of containation of various sources of drinkingwater with cryptosporidium and effects on people with and without HIV infection.
Design: Prospective, Investigative study.
Methods: Water samples, from Highly and lowly populated site areas were
surged through filters using controlled cavity metering pump. filters were teased
into 0.01% Poly Oxy ethylene Sorbitan mono oleate solution processed for
smears. Adults (some with HIV and others without) and children's (All without HIV
infection) faeces were also processed for smears. All the smears were analysed
by Modifield Ziehl-Neelsen and flourescent Acridine methods. The gold standard
for cryposporidium was the one positive with both methods.
Results: A total of 100 water samples analysed, 56% (56/100) had oocysts
of cryptosporidium in them. Other pathogenic contaminants accounted for 2%
(2/100). Adults, 32 in number, 72% (23/23) with HIV infection and cryptosporidium
oocysts in their faeces had continous diarrhoea (cryptosporidiosis) of more than
1 month. In all the HIV negative 20 children, 60% (12/12) below the age of 2
years had mild diarrhoea for not more than 8 days, while those above 2 years
were assymptomatic.
Conclusion: Cryptosporidium is a definite pathogen in Drinking-water and
poses a danger of life-threating chronic diarrhoea to immune compromised patients like those with HIV infection and AIDS. Children below the age of 2 years
are at a risk of contracting cryptosporidiosis when exposed to parasites.
| 32101 | High detection rate of HGV in blood plasma, but not
seminal fluid plasma of HIV-1 infected individuals
Graeme J. Moyle1, Jonathan K. Ball2, M. Crowe2, R.C. Hollingsworth2,
W.L. Irving2, M. Youle3, D. Pillay4. 1 Chelsea and Westminster Hospital, 369
Fulham Road, London, SWIO 9NH; 2University of Nottingham, Nottingham;
3Royal Free Hospital, London; 4Heartlands Hospital, Birmingham, UK
Background: The incidence of Hepatitis G (HGV) amongst people with HIV is
not currently known. Precise routes of transmission for HGV are currently under
evaluation.

Methods: Fifty paired seminal fluid and blood samples were obtained from
a cohort of HIV-1 infected homosexual men individuals in whom there was no
evidence of parenteral exposure to infectious agents. RNA was extracted using
components from the HIV-1 quantitative NASBA/Nuclisens assay and tested for
the presence of HGV RNA using primers complementary to conserved regions
of the 5' NCR. HIV-1 RNA was quantified using either the NASBA or Nuclisens
assays