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Background During the 2017/18 and 2018/19 influenza seasons, molecular amplification-based point-of-care tests (mPOCT) were introduced in Scotland to aid triaging respiratory patients for hospital admission, yet communication of results to national surveillance was unaccounted for. Aim This retrospective study aims to describe steps taken to capture mPOCT data and assess impact on influenza surveillance. Methods Questionnaires determined mPOCT usage in 2017/18 and 2018/19. Searches of the Electronic Communication of Surveillance in Scotland (ECOSS) database were performed and compared with information stored in laboratory information management systems. Effect of incomplete data on surveillance was determined by comparing routine against enhanced data and assessing changes in influenza activity levels determined by the moving epidemic method. Results The number of areas employing mPOCT increased over the two seasons (6/14 in 2017/18 and 8/14 in 2018/19). Analysis of a small number of areas (n = 3) showed capture of positive mPOCT results in ECOSS improved between seasons and remained high (> 94%). However, capture of negative results was incomplete. Despite small discrepancies in weekly activity assessments, routine data were able to identify trend, start, peak and end of both influenza seasons. Conclusion This study has shown an improvement in capture of data from influenza mPOCT and has highlighted issues that need to be addressed for results to be accurately captured in national surveillance. With the clear benefit to patient management we suggest careful consideration should be given to the connectivity aspects of the technology in order to ensure minimal impact on national surveillance.

Background This report describes the investigation and public health management of a community-based outbreak of severe adenovirus serotype 14p1 respiratory infection affecting the Tayside area during 2011. It is the first report of an adenovirus outbreak involving prisons. Methods An outbreak-based/incident management approach was carried out. Alerts were sent out to local doctors, general practitioners, prison healthcare staff and consultants so that cases could be identified prospectively. Sequencing of hexon, fibre and E1A regions of adenovirus were carried out to genotype the viruses. Results Fifteen cases were identified in total, including 13 confirmed cases and 2 possible cases. There were 3 deaths amongst the 13 confirmed cases, with a case fatality rate of 23%. Eight of the cases had a direct association with one of the two prisons in the area. Conclusions We advise that surveillance measures for adenovirus infection and guidelines for the management of critically ill patients should be developed in order to identify outbreaks at an early stage and allow patients to receive appropriate treatment. Adenovirus infection should be borne in mind as a cause of severe pneumonia in closed settings such as prisons.

Nucleic acid amplification tests (NAATs) are the most sensitive method for diagnosing chlamydia and gonorrhoea. We use the COBAS 4800 CT/NG combined assay (Roche Molecular Diagnostics, CA, USA), and whilst the majority of samples yield definitive results, a small proportion are reported as indeterminate. In these instances, it is usual practice to request repeat samples which delays diagnosis. This audit was twofold: first to establish the proportion of indeterminate results with current NAAT testing requiring re-sampling. Second, to determine whether a second NAAT such as Cepheid GeneXpert CT/NG assay (Cepheid, CA, USA) could be used on initial indeterminate samples to resolve indeterminate results, therefore reducing need for repeat sampling. During 2012, 144/21,931 (0.66%) samples were indeterminate for Neisseria gonorrhoeae, Chlamydia trachomatis or both, and a repeat sample was received in only 51.77% of patients with final results being delayed for more than 24 h. Over the next six months, there were 77/9472 (0.81%) indeterminate results. After an evaluation and introduction of the Cepheid assay, the number of indeterminate results fell to 9 (0.10%). Thus, use of the Cepheid assay significantly reduced indeterminate results, reduced reliance on a repeat sampling and significantly improved turnaround time, laboratory workflow and patient experience.

Objectives: To describe the resistance mutations selected by a first-line regimen of zidovudine/lamivudine/tenofovir in the absence of real-time viral load monitoring. Design: A substudy of 300 participants from the Development of Antiretroviral Therapy in Africa trial in Uganda and Zimbabwe, which compared managing antiretroviral therapy with and without laboratory monitoring. Methods: Stored plasma samples from selected time points were assayed retrospectively for HIV-1 RNA. The pol gene in all baseline samples and those with HIV RNA >1000 copies per milliliter at weeks 24 and 48 were sequenced. Results: The proportion with HIV RNA >1000 copies per milliliter increased from 15% at 24 weeks to 24% at 48 weeks. Eighteen of 31 (58%) genotyped samples at 24 weeks had ≥1 major nucleoside reverse transcriptase inhibitor-associated mutations compared with 41 of 47 (87%) at 48 weeks. Excluding 1 nonadherent patient, a mean of 2.0 (95% confidence interval: 1.3 to 2.8) thymidine analogue mutations (TAMs) developed between weeks 24 and 48 among 14 patients with HIV RNA >1000 copies per milliliter at both time points. K65R was detected in 8 of 63 (13%) patients and was negatively associated with number of TAMs (P = 0.01) but not viral subtype (P = 0.30). Conclusions: A high rate of acquisition of TAMs, but not of K65R, among patients with prolonged viraemia was observed. However, most patients were virologically suppressed at 48 weeks, and long-term clinical and immunological outcomes in the Development of Antiretroviral Therapy in Africa trial were favorable.

Humoral responses against extra-cellular HIV-1 Tat may be beneficial as Tat has been implicated in the viral pathogenesis associated with HIV-1 disease progression. We determined the levels of anti-Tat IgG in sera of HIV-1 seropositive individuals from the Rural Clinical Cohort in Uganda using nine different Tat proteins representative of the major subtypes presently accounting for 97% of infections worldwide. We observed the presence of anti-Tat IgG able to react against the various subtypes tested, although none cross-reacted against all nine variants. We show that 46.25% of seropositive patients were able to recognise at least one Tat variant with 1:1000 sera dilution. We also show that the C terminus of Tat is the most variable region and an important epitope that might explain the limitation of cross-recognition of Tat antibodies regarding Tat variants. This study shows in seropositive patients that Tat can tolerate mutations without modification of its primary function but with changes in its immunogenic properties. These findings should be considered when designing Tat-based HIV-1 vaccines.

Antigenic properties of the V3 region are reflected by HIV-1 serotypes. These may represent biological properties of the virus. We serotyped HIV-1 in 142 serum samples from participants in a rural Uganda cohort who seroconverted between August 1991 and December 2001. Clinical progression was assessed using Cox proportional hazards and Kaplan-Meier methods. Of 112 (79%) samples successfully serotyped, 36% were serotype A, 17% serotype B, 18% serotype C, and 29% serotype D. Median follow-up time, age at enrollment, and first CD4 count were similar in each serotype group. Clinical progression was faster for serotype D than other serotypes to AIDS or death, death, and CD4 count

International audience; AIDS vaccines now use a truncated version of 86 residues of the Tat protein related to the HIV-1 HXB2 strain predominant in Europe and North America. We compared antibodies raised in rabbits using a B subtype short Tat HXB2(86) and a full-length Tat HXB2(100). Serum against HXB2(86) recognizes only B and D subtypes while serum against HXB2(100) recognizes B, D, and C subtype variants. Conformational epitopes appear to be involved in the capacity of anti-Tat HXB2 sera to recognized non-homologous Tat variants. A linear B-epitope identified in sequence 71-81 in HXB2(86) disappears in HXB2(100), which has a new linear B-epitope identified at the C-terminus. Anti-HXB2(100) serum has a higher titer in neutralizing antibody against homologous and non-homologous variants compared to anti-HXB2(86) serum. We suggest that a Tat vaccine should contain a Tat variant with regular size, up to 99-101 residues now found in the field.

Investigations concerning the extent and nature of subtype-specific and intersubtype immune responses in HIV-1-infected persons are necessary for the development of appropriate candidate vaccines. In the cross-sectional study described here, 26 HIV-1-positive Ugandan patients were tested for their ability to mount HIV antigen-specific cellular immune responses. Subjects were infected with either HIV-1 subtypes A, C, or D. Recombinant vaccinia virus (rVV)-based and peptide-based enzyme-linked immunospot (Elispot) assays were used to evaluate HIV-1-specific gamma-interferon (IFN-gamma) cellular responses. rVV expressing gag, pol, or env proteins derived from HIV-1 subtypes A, B, and D were evaluated for their ability to induce whole HIV-1-protein-specific IFN-gamma responses in 14 patients. A panel of previously identified HLA class I-restricted peptides based on representative sequences from HIV-1 subtypes A, B, C, and D and restricted through HLA-A2, -A29, -B42, -B53, and -B57 alleles were used to evaluate the presence of HIV-1-peptide-specific T cells in 19 patients. Using rVV, 27 of a possible 38 subtype-specific responses (71%) and 56 of a possible 110 intersubtype responses (51%) were observed. When appropriate peptides were used 18 of 39 (46.2%) subtype-specific and 13 of 39 (33.3%) intersubtype responses were observed. Peptide responses were higher quantitatively than those seen when rVV were used. In 7 patients, both rVV and specific peptides were evaluated; in 3 of 7 individuals, global responses were seen despite a lack of measurable HLA-restricted peptide-specific responses demonstrating the need to evaluate a broader range of HIV-specific immune responses.

CD4(+) T cell help is important for the functionality of CD8(+) cytotoxic T-lymphocytes (CTLs) in limiting viral replication and may contribute to mediation of apparent resistance to HIV-1 infection in exposed seronegative (ESN) individuals. Using five HIV-1 antigens in an intracellular cytokine assay, the presence of specific antigen-responsive interferon- gamma-positive (IFN-gamma(+)) CD69(+) CD4(+) T-lymphocytes was evaluated in ESNs, their seropositive partners, and unexposed seronegative controls. Ten ESNs (five females, five uncircumcised males) were identified from 10 HIV-1 serodiscordant couples with a history of frequent unprotected sexual intercourse. All ESNs and controls were negative on two EIAs and for HIV-1 proviral DNA. The frequency of ESNs with antigen-responsive IFN-gamma(+) CD69(+) CD4(+) T-lymphocytes ranged from three to five of eight for the different HIV-1 antigens. Six of eight ESNs tested had a positive response to at least one of the five antigens. Responses were on average 3.5 times higher among seropositives compared to ESNs and absent in the five unexposed controls. A negative correlation was noted between responses in ESNs and the plasma viral load of their seropositive spouse. Clade-specific and cross-clade reactivity were noted in both ESNs and seropositive partners tested. The findings confirm that ESNs are in a state of HIV-1-specific immune activation and suggest that HIV-1-specific IFN-gamma(+) CD69(+) CD4(+) T-lymphocytes in addition to HIV-1-specific CD8(+) CTLs already described by others are potential immunological correlates of protection from persistent HIV-1 infection.

The effect of human immunodeficiency virus (HIV) type 1 envelope subtypes A and D on disease progression was investigated in 1045 adults in Uganda. At enrollment and every 6 months, a clinical history, examination, and laboratory investigations that included CD4 cell counts were done. HIV-1 envelope subtype was assessed mainly by peptide serology supplemented by heteroduplex mobility assay and DNA sequencing. A multivariate analysis of survival was performed to assess the prognostic value of HIV-1 subtype on death. A marginal general linear model also determined the effect of subtype on CD4 cell count during follow-up. Subtype D was associated with faster progression to death (relative risk, 1.29; 95% confidence interval, 1.07-1.56; P=.009) and with a lower CD4 cell count during follow-up (P=.001), compared with subtype A, after adjusting for CD4 cell count at enrollment. In Africa, envelope subtype D is associated with faster disease progression, compared with subtype A.

We describe the burden of influenza B infections in Scotland during a 13-year study period. Influenza A and B viruses cocirculated throughout the period, with numbers of influenza B cases approaching or exceeding those of influenza A during six influenza seasons. Influenza B viruses of both Victoria and Yamagata lineage were detected in two of six seasons investigated. For the 2012/13 season, influenza B accounted for 44.4% of all influenzas, with the highest incidence in those under the age of five years. Influenza B virus infections led to fewer admissions to an intensive care unit (ICU) and a lower mortality rate than influenza A (37 vs 81 ICU admissions and three vs 29 deaths) during the 2012/13 season. However, a quarter of those admitted to ICU with influenza B had not been immunised and 60% had not received specific influenza antiviral therapy. This highlights the need for consistent influenza vaccination and prompt usage of antiviral treatment for identified risk groups. Combining the newly introduced vaccination programme for children with the use of a tetravalent vaccine may provide the opportunity to improve the control of influenza B in those with the highest influenza B burden, children and young adolescents.

We compared HIV-1 RNA levels using the nucleic acid sequenced based amplification (NASBA) test kit in 2 samples taken one year apart from participants infected with env subtype A or D in a population-based cohort in Uganda. Fifty participants were infected with subtype A and 70 with subtype D. HIV-1 RNA levels were significantly higher in subtype D unadjusted ( P = 0.001), and after adjusting for age, gender, and CD4 count ( P < 0.001). Eighty-six participants had HIV-1 RNA measurements in both years and 67 (78%) were within one log10 of their result a year before. There was no relationship between the difference in log viral load and proportion of CD4 change. Individuals infected with subtype D had a higher average increase in viral load and this was statistically significant if adjusted for baseline levels and CD4 count ( P = 0.015).

The study was based in south-west Uganda where significant differences in HIV prevalence have been found between urban and rural areas. Longitudinal data collected in a diary format was used to determine the extent to which high-risk men and women living in a truck stop/trading town had sexual contact with people from surrounding rural areas and a nearby fishing village. Study participants were 143 men, 75 of whom were resident in the town, 40 in a fishing village and 28 in rural areas, and 81 women, of whom 47 were resident in the town, 25 in the fishing village and 9 in a rural area. During 1687 man weeks the 143 men made 3149 trips and had 5189 sexual contacts. Ninety-two per cent of these sexual contacts occurred in the man's current place of residence and 21% were with a new partner. The 81 women participated for 1280 women weeks during which they recorded 6378 sexual contacts. Women who lived in the fishing village and the rural area had around 90% of their contacts with local men while those who lived in the town fell into 3 categories: women who charged a relatively high price for commercial sex had only 11% of contacts with men living in the town, while those who charged a tenth of the price had 71% of contacts with town men. The small number of women who fell into an intermediate category, in terms of price, had sexual contact with a wide variety of men. These findings show that there is little scope for HIV infection to spread between different residential or occupational groups. This may help to explain how large differences in HIV seropositivity between neighbouring localities can be maintained for long periods, despite considerable social and economic mixing between groups and high levels of sexual partner change within groups.In southwest Uganda, there is concern that sexual mixing between rural areas with low HIV prevalence and urban areas with high HIV prevalence will produce uniformly high rates of HIV. To determine the extent of such mixing, a prospective study was conducted in a trading town on the trans-African highway with a steady flow of male truckers, an agricultural hinterland to the west, and a fishing village to the east. A total of 143 men recruited largely from bars and 81 women reported to be sexually promiscuous kept records of their sexual contacts over a 6-month period. During 1687 man-weeks of observation, the men made 3149 road trips and had 5189 sexual contacts; 92% of these contacts occurred in the men's place of residence and 21% involved a new partner. 59% of town men's sexual contacts, 61% reported by men in the fishing village, and 52% of those in rural areas involved casual partners. An additional 6378 sexual encounters were recorded during 1280 woman-weeks. Close to 90% of women's sexual contacts in the 2 rural areas involved local men. Among the highest-paid town commercial sex workers, only 11% of sexual contacts involved men from the town; in contrast, 71% of sexual encounters among town women who charged only 10% the price of their more expensive counterparts were with local men. Serologic testing of a subset of 75 men and 52 women yielded HIV prevalences of 28% and 52%, respectively. The distinct sexual networks identified in this study suggest it is unlikely that rates of HIV infection in the rural areas will reach those in nearby trading towns.

We investigated the feasibility of using vaccinia virus (VAC) recombinants containing large multigene fragments of orf virus DNA to identify protective antigens of orf virus (OV). Sixteen OV strain NZ2 DNA fragments with an average size of 11.4 kb were recombined into VAC strain Lister. Each fragment was mapped relative to OV restriction endonuclease maps but was otherwise uncharacterized. Together the recombinants represent 95% of the OV genome in an overlapping manner. Immunofluorescence showed all 16 constructs expressed products recognized by OV antiserum and radioimmune precipitation with the same antiserum allowed the localization of the major antigens of OV to specific recombinants. These data indicated the approximate genomic locations of the genes encoding the OV major antigens and showed that their expression was authentic rather than resulting from read through from VAC sequences adjacent to the site of recombination. Vaccination of OV-naive sheep with the recombinant library provided protection against a subsequent challenge with virulent OV. These data confirm the feasibility of the proposed strategy.

The association of tuberculosis (TB) with HIV infection is well established with TB being one of the common manifestations of AIDS. The proportion of TB cases with HIV infection has been rising and the global prevalence of TB/HIV coinfection among adults aged 15–49 years is 0.36%. India has the largest number of co-infected individuals. Bangladesh ranks fifth among the countries with the highest burden of TB but fortunately HIV prevalence in Bangladesh is still low. However recent data from surveillance show that Bangladesh is at the brink of a concentrated epidemic in injecting drug users. The country has 363 people with HIV/AIDS (PHA) identified through passive case reporting as announced by the National AIDS/STD Programme of the Government of Bangladesh on 1 December 2003. Of 67 PHA who visited the Voluntary Counselling and Testing (VCT) Unit of ICDDRB between April 2003 and June 2004 47 had opportunistic infections of whom six had TB (M M Parvez ICDDRB personal communication). (excerpt)

Following orf virus reinfection in sheep, CD8+ T cells were recruited to the lesion site in the skin during the period of viral replication and appeared in increased numbers in afferent lymph draining the infection site. A proportion of the CD8+ T cells were activated, particularly in the skin, as determined by their expression of the cytolytic cell-associated serine protease, BLT-esterase. This was a useful marker for activated cytolytic cells as there was a good correlation between afferent lymph CD8+ T cell-mediated lysis of autologous orf virus-infected skin cells and their expression of BLT-esterase in vitro. The majority of the cells that expressed BLT-esterase in the skin and lymph were CD8+ T cells and cytolysis in vitro was predominately by the MHC class I antigen presentation pathway. The conclusion is that orf virus reinfection is characterized by limited viral replication and stimulation of an activated CTL response in the skin.

Background Boosted protease inhibitor (bPI) monotherapy (bPImono) potentially has substantial cost, safety and operational benefits. It has never been evaluated as second-line antiretroviral therapy (ART) in Africa. Methods After 24 weeks of lopinavir/ritonavir- containing second-line therapy, DART participants were randomized to remain on combination therapy (CT), or change to bPI-mono maintenance (SARA trial; ISRCTN53817258). Joint primary end points were CD4+ T-cell changes 24 weeks later and serious adverse events (SAEs); retrospectively assayed viral load (VL) was a secondary end point. Analyses were intention-to-treat. Results A total of 192 participants were randomized to CT ( n=95) or bPImono ( n=97) and followed for median 60 weeks (IQR 45–84). Participants received median 4.0 years (IQR 3.5–4.4) first-line ART. Median CD4+ T-cell count at first-line failure was 86 cells/mm3 (47–136), increasing to 245 cells/mm3 (173–325) after 24-week induction when 77% had VL+ T-cell increase was 42 (CT, n=85) versus 49 cells/mm3 (bPImono, n=88; adjusted difference 13 [95% CI -15, 43], P=0.37; non-inferior compared with predetermined non-inferiority margin [-33]). Virological suppression was greater for CT versus bPImono (trend P=0.009): 77% (70/91) versus 60% (56/94) were 24 weeks after randomization, and 5 (2 CT and 3 bPImono) experienced SAEs ( P=0.51). Conclusions bPImono following a 24-week second-line induction was associated with similar CD4+ T-cell response, but increased low-level viraemia, generally without protease inhibitor resistance. Longer-term trials are needed to provide definitive evidence about effectiveness in Africa.

In common with reports from other European countries, we describe a substantial increase in the number of laboratory reports of Mycoplasma pneumoniae in Scotland in 2010 and 2011. The highest number of reports came from those aged one year and younger. However, reports from young children were more likely to come from PCR testing than serological testing.

Hysterectomy-procured, barrier maintained lambs were immunised with either orf virus or vaccinia virus and subsequently challenged with both viruses. Under these conditions lambs were protected from challenge with the homologous virus but no cross-protection was observed. The feeding of colostrum that contained antibodies to orf virus had no effect on the duration of viral lesions. Immunoblotting analysis and ELISA of serum samples taken during the course of the experiment indicated that the animals mounted antibody responses to both viruses. The cross recognition of 3 vaccinia virus antigens by the hyperimmune anti-orf virus serum was revealed by immunoblotting.

Injecting drug users (IDU) were enrolled from two detoxification clinics and two needle/syringe exchange programmes (NEP) in central and northwest Bangladesh. Syphilis, hepatitis C and HIV rates were highest in IDU from the NEP of central Bangladesh (23, 66.5 and 1.4%, respectively), whereas current hepatitis B infection rates were highest in IDU from the NEP of northwest Bangladesh (12%). Five HIV strains were subtype C and one E/B. The 32 base pair (bp) deletion of the CCR5 gene was not detected.

BACKGROUND: We investigated virological response and the emergence of resistance in the Nevirapine or Abacavir (NORA) substudy of the Development of Antiretroviral Treatment in Africa (DART) trial. METHODS: Six hundred symptomatic antiretroviral-naive human immunodeficiency virus (HIV)-infected adults (CD4 cell count, 1000 copies/mL, the residual activity of therapy was calculated as the reduction in HIV RNA level, compared with baseline. RESULTS: Overall, HIV RNA levels were lower in the nevirapine group than in the abacavir group at 24 and 48 weeks (P < .001), although no differences were observed at weeks 4 and 12. Virological responses were similar in the 2 treatment groups for baseline HIV RNA level

— Following dermal infection of previously sensitized sheep with orf virus, differences in cell phenotype and kinetics are observed in afferent lymph flowing into the local lymph node compared to efferent lymph exiting the node. Afferent lymphocytes in five out of six sheep synthesized IFN-γ (an indicator of T-cell activation) 10–30 h. after challenge, whereas none was synthesised by efferent lymphocytes until 4–11 days after infection when production was intermittent and variable. Both afferent (five out of six) and efferent (six out of seven) lymph plasmas contained IFN-γ at 10–30 h. but none was found in efferent lymph after 30 h. Colony stimulating activity (an indicator of immunoinflammatory responses) in both lymph compartments was biphasic peaking at 10–30 h. with a later peak which coincided with a lymphoblast response. The results are consistent with an early memory T-cell response in the dermis and afferent lymph followed by recruitment of antigen-specific lymphocytes from the blood, and activation within the node. Resume— Apres infection dermique de noutons prealablement sensibilises ail virus de l'ecthyma contagieux, des differences de phenotype cellulaire et de cinetique ont ete observes entre le flux lymphotique afferent au ganglion local et la lymphe quittant le ganglion. Les lymphocytes afferents chez cinq des six moutons synthetisaient de l'IFNγa 10–30 h mais aucun n'a ete retrouve dans la lymphe efferente a 30 h. L'activite de stimulation de colonie (un indicateur des reponses immunoinflammatoires) dans les deux compartiments lymphatiques suivait deux pics a 10 et 30 h avec un dernier pic qui coincidait avec la reponse lymphoblastique. Les resultats sont compatibles avec une reponse precoce de cellules T memoire dans le derme et les lymphatiques afferents suivie par le recrutement de lymphocytes specifiques de l'antigene dans le sang, et leur activation dans les ganglions. Zusammenfassung— Nach einer dermalen Infektion von vorber sensibilisierten Schafen mit Orf Virus konnten Unterschiede im Zell Phanotyp und in der Kinetik der afferenten Lymphe (in den ortlichen Lymphknoten fliesend) gegenuber der efferenten Lymphe (aus dem Lymphknoten abfliesend) gemessen werden. Die afferenten Lymphozyten synthetisierten bei 5 von 6 Schafen IFN-y (ein Indikator der T-Zell Aktivierung) 10 bis 30 Stunden nach der Infektion, wahrend keines von der efferenten Lymphozyten synthetisiert wurde; erst 4 bis 11 Tage nach der Infektion zeigte sich eine intermittierende und variable Produktion. Sowohl bie 5 von 6 afferenten wie bei 6 von 7 efferenten Proben enthielt das Lymphplasma IFN-y nach 10 bis 30 Studen, aber keines in den efferenten Proben nach 30 Studen. Die “Kolonie-stimulierende Akrivitat” als ein Indikator fur Immuminflammatorische Reaktionen zeigte in beiden Lympharten einen zweiphasigen Peak nach 10 bis 30 Stunden, wobei der spatere Peak mit der Lymphoblastenreaktion zusdammen fiel. Diese Ergebnisse sprechen fur eine fruhe Reaktion der Memory-T-Zellen in der Dermis und der afferenten Lymphe mit anschliesender Rekrutierung Antigen-spezifischer Lymphozyten aus dem Blut und Aktivierung innerhalb des Lymphknotens. Resumen Despues de la infeccion dermica de ovejas previamente sensibilizadas con el virus Orf, se obsevaron las diferencias en el fenotipo y quinetica celular de de los nodolus aferentes que desembocaban en los nodulos locales, comparados con los de tipo eferente. Los linfocitos aferentes de 5/6 ovejas sintetizaron IFN-y, (un indicador de la activacion de los linfocitos T), despues de 10–30 horas, pero no se encontro alguno en los linfocitos eferentes hasta los dias 4–11 despues de la infeccion, cuando la produccion era intermitente y variable. Ambos plasmas linfociticos, aferente (5/6) y eferente (6/7), contenian IFN-y a las 10–30 horas, pero no se encontro alguno en los eferentes hasta despues de las 30 horas. En ambos compartimentos linfaticos la actividad estimulante de la colonia, (un indicador de respuesta inmunoinflamatoria), presento un maximo de tipo bifasico a las 10–30 horas con otro maximo tardio, que coincidio con la respuesta de tipo linfoblastico. Los resultados corresponden con una respuesta de los linfocitos T de memora temprana en la dermis y los nodulos linfaticos aferentes, seguidos por un flujo de linfocitos antigeno-especificos procedentes de la sangre y activacion dentro del nodulo linfatico.

BACKGROUND: Some HIV infected individuals remain asymptomatic for protracted periods of time in the absence of antiretroviral therapy (ART). Virological control, CD4 T cell loss and HIV-specific responses are some of the key interrelated determinants of HIV-1 disease progression. In this study, possible interactions between viral load, CD4 T cell slopes, host genetics and HIV-specific IFN-gamma responses were evaluated in chronically HIV-1-infected adults. METHODOLOGY/PRINCIPAL FINDINGS: Multilevel regression modeling was used to stratify clade A or D HIV-infected individuals into disease progression groups based on CD4 T cell slopes. ELISpot assays were used to quantify the frequency and magnitude of HIV-induced IFN-gamma responses in 7 defined rapid progressors (RPs) and 14 defined slow progressors (SPs) at a single time point. HLA typing was performed using reference strand conformational analysis (RSCA). Although neither the breadth nor the magnitude of the proteome-wide HIV-specific IFN-gamma response correlated with viral load, slow disease progression was associated with over-representation of host immunogenetic protective HLA B* alleles (10 of 14 SPs compared to 0 of 7; p = 0.004, Fisher's Exact) especially B*57 and B*5801, multiclade Gag T-cell targeting (71%, 10 of 14 SPs compared to 14%, 1 of 7 RPs); p = 0.029, Fisher's Exact test and evident virological control (3.65 compared to 5.46 log10 copies/mL in SPs and RPs respectively); p

— Changes in cell output and phenotype were monitored in the afferent lymph draining the cutaneous site of orf virus infection. Total cell output was increased although, relative to mock inoculation, output of non-specific esterase positive (NSE) cells (macrophage/monocyte) was reduced. In contrast, the proportion of acetylcholine esterase positive (Ache) cells (erstwhile epidermal Langerhans' cells) in the afferent lymph was increased in response to viral challenge. Afferent lymph cells demonstrated an increase in spontaneous and antigen stimulated in vitro lymphoproliferation. However, in some infected animals, a temporary period of reduced lymphoproliferation to exogenous orf virus antigen (but not nematode antigen) was recorded. Other changes specific to viral challenge were the appearance of polymorphonuclear cells (PMNs) and blast cells in the afferent lymph after 24 hours and five days respectively. The relevance of these changes with respect to control of orf virus infection is discussed. Resume— Les modifications de debit cellulaire et de phenotype de la lymphe drainant les sites d'infection par le virus de la dermite pustuleuse contagieuse ovine oht ete mesurees. Le nombre total de cellules drainees etait augmente, bien que compare aux inoculations experimentales, le debit de cellules esterase non specifique positives (NSE) (macrophages/monocytes) etait diminue. Par contre, la proportion de cellules acethyl choline esterase positioves (AChe) (cellules de Langerhans) dans la lymphe afferente etait augmente apres une inoculation virale. Les cellules de la lymphe afferente presentaient une augmetation de la reponse lymphoproliferative in vitro spontannee et induite par l'antigene. Toutefois, chez certains animaux, une periode transitoire de diminution de la response lymphoproliferative a des antigenes de virus de la dermite pustuleuse contagieuse ovine exogenes (mais pas aux antigenes de Nematodes) a ete notee. Les autres modifications specifiques de l'inoculation virale etaient l'apparition de polynuleaires (PMNs) et de cellules blastiques dans le lymphe aferente au bout respectivement de 24 heures et cinq jours. La signification de ces modifications par rapport aux temoins est discutee. Zusammenfassung— In der afferenten Lymphe, die Hautbezirke mit Orf-Virusinfektion drainierte, wurden Veranderungen im Zellgehalt und Zellphanotyp registriert. Der Gesamtaussetos an Zellen war erhoht, obwohl hinsichtilich einer Vergleichsinokulation der Ausstos an nicht-spezifischen Esterase-positiven (NSE) Zellen (Makorphagen/Monozyten) vermindert war. Im Gegensatz dazu waren die Azetylcholinesterase-positiven (AChe) Zellen (ehemals epidermale Langerhans Zellen) in der afferenten Lymphe vermehrt anzutreffen als Reaktion auf die virale Herausforderung. Die Zellen in der afferenten Lymphte zeigten in vitro einen Anstieg in der spontanen und antigenstimulierten Lymphoproliferation. Es wurde jedoch bei einigen betroffenen Tieren auch uber eine temporare Phase mit verminderter Lymphoproliferation auf exogenes Orf-Virusantigen (nicht aber auf Nematodenantigen) berichtet. Andere, spezifisch virusinduzierte Veranderungen bestanden im Auftreten polymorphonuklearer Zellen (PMNs) und Blastzellen in der afferenten Lymphe 24 Stunden bzw. funf Tage nach der Infektion. Die Relevanz dieser Veranderungen unter Berucksichtigung der Bekampfung von Orf Virusinfektionen wird diskultiert. Resumen Los cambios en el rendimiento y el fenotipo celular fueron monitorizados en los linfaticos aferentes que irrigaban un punto en la piel, que a su vez padicia de infeccion virica con el virus orf. El rendimiento celular total se vio aumentado, aunque con respecto a una inoculacion simulada, el rendimiento de celulas no-especificas esterasa positiveo (NSE), se vio reducida (macrofagos/monocitos). En cambio, la proporcion de celulas acetilcolina esterasa positivo, (AChe) (anteriormente llamadas celulas epidermicas de Langerhans) en el linfatico aferente, aumento en respuesta a la infeccion virica. Las celulas del linfatico aferente demonstraron un incremento en linfoproliferacion espontanea y linfoproliferacion estimulada antigenicamente in vitro. Sin embargo, en algunos de los animales infectados, se observo un periodo temporal de linfoproliferacion reducida hacia el antigeno del virus orf (pero no al antigeno del nematodo). Otros cambios especificos al desafio virico feuron la aparicion de polimorfonucleares (PMNs) y blastocitos en los linfaticos aferentes despues de 24 horas y 5 dias respectivamente. La importancia de estos cambios con respecto al control del virus orf es discutida.

There are data to suggest that both the humoral and cellular immune responses directed against Tat are beneficial in delaying HIV disease progression. We examined the association between the occurrence of Tat-specific binding antibodies (Abs) and different parameters of HIV-1 disease progression. We generated eight Tat proteins, derived from HIV-1 subtypes A, B, C, and D, and circulating recombinant form CRF01_AE. These proteins were used to screen for Tat-specific binding Abs by an ELISA. Using five Tat proteins, we investigated whether the occurrence of Tat-specific Abs within 2 years after seroconversion for the majority, affected disease progression over time among 126 participants using survival analysis and rate of CD4 decline. Of these, 52 participants with a sample at 1.5 and 4.5 years after seroconversion were further examined to study the effect of Tat-specific Ab loss or maintenance on disease progression. Finally, using all the eight Tat proteins, we also investigated whether specific Abs to these Tat proteins among 48 participants, grouped as rapid progressors (RP, n = 26) and long-term survivors (LTS, n = 22) according to their CD4 decline over time, affected disease progression. Survival analysis did not reveal any evidence of protection from progression by Tat-specific Abs. Comparison of rate of CD4 declines between individuals with and without Abs to any Tat protein showed only a small and borderline significant advantage of having Tat-specific Abs (p = 0.043). There was no correlation between either loss or maintenance of Tat-specific Abs and disease progression. Comparison of LTS with RP showed no evidence that Tat-specific Abs slows participants' disease progression. This study showed no evidence of a protective effect of having Tat-specific Abs among these Ugandan subjects.

SUMMARYThe purpose of this study was to document the dynamics of HIV-1 subtypes in Scotland over a 6-year period. Viral RNA from all-new diagnoses was amplified by nested PCR and sequenced in the gag and/or env regions. Subtype was assigned by phylogenetic analysis, and aligned with demographic data including likely route and geographical origin of infection. We present data on 80% of all new diagnoses in Scotland between April 2000 and April 2006. Within the background of an expanding epidemic, subtype B predominates in men who have sex with men and intravenous drug users but there is a small but consistent number of UK-acquired infections in these risk groups caused by non-B subtypes. In heterosexuals, non-B subtypes acquired abroad, especially Africa, are still the largest group but again UK-acquired numbers are rising. The social and clinical significance of the spread of non-B subtypes in different ethnic and risk groups remains to be established.

Although not linked to a disease, GB virus-C viraemia has been associated with an improved prognosis in HIV-1-co-infected individuals. Most studies have been conducted on men (men who have sex with men or injection drug users) infected with HIV-1 subtype B, whereas here we report on both male and female subjects from rural Uganda, predominantly infected via the heterosexual route with HIV-1 subtypes A and D. In a longitudinal study of 272 participants, 47 were GBV-C positive and 181 negative, as determined by reverse transcription-polymerase chain reaction, in both of two plasma samples taken a median of 5.0 years apart. The remainder either acquired (25) or cleared (19) infection. Multilevel regression analyses and Cox survival analyses revealed that participants chronically infected with GBV-C had a slower decline in CD4+ T cells ( P

The objectives were to determine whether there are differences in coreceptor use in subjects infected with HIV-1 envelope subtypes A and D that could explain the differences in progression rates between these subtypes in a rural Ugandan cohort. HIV-1 was subtyped in env by V3 sequencing or heteroduplex mobility assay. Coreceptor use was determined by the ability of the isolates to replicate in U87 CD4+ cells expressing different coreceptors. The Fisher exact test was used to examine the relation between coreceptor use and subtype clinical stage and V3 charge. The Kruskall-Wallis nonparametric test was used to examine the association between median CD4 cell counts coreceptor use and subtype. Logistic regression was used to examine predicted coreceptor use at different CD4 groupings. Isolates from 66 participants were analyzed. Thirty-one were infected with subtype A and 35 were infected with subtype D. Although this work was based on a small sample size we found statistically significant differences. The probability of having an X4 virus was higher in subtype D infections than in subtype A infections among those with a non-AIDS clinical status (Fisher exact test P = 0.040). Logistic regression analysis in which we predicted X4 use by subtype and stratified by CD4 group confirmed these findings among those with a CD4 count > 200 cells/µL (likelihood ratio test P = 0.003). R5 viruses were associated with higher median CD4 cell counts than X4 or X4/R5 (Kruskall-Wallis test P = 0.0045). AV3 charge of +5 and greater was highly associated with X4 virus (Fisher exact test P = 0.006). These subtype differences in coreceptor use may partially explain the faster progression rates we have previously reported in individuals infected with subtype D compared with subtype A. Our observations may have implications for the future use of coreceptor inhibitors in this population. (authors)

The apparent natural transmission of orf virus from clinically normal ewes to susceptible sheep was observed during a border disease vaccine experiment. The 14 susceptible sheep were persistently infected with border disease virus and had been reared indoors in isolation from other sheep since birth. Their ages ranged from two to four years and they were housed in two groups; group 1 consisted of four sheep persistently infected with the Moredun strain of border disease virus and group 2 consisted of 10 sheep persistently infected with the Oban strain of the virus. On day 0, six sheep were removed from group 2 and rehoused. To the remaining four sheep in each group were added eight four- to six-year-old pregnant conventionally reared ewes at 48 days gestation. Fourteen days later the four sheep in group 1 were moved to another pen housing eight similar five-year-old pregnant ewes at 48 days' gestation, and the four sheep from group 2 were rehoused with their original stallmates. Twenty-one days later lip lesions typical of orf were first observed on the sheep from both groups and the disease spread to all the sheep persistently infected with border disease virus over the next four weeks. Virological and serological evidence demonstrated that the source of infection for the sheep was almost certainly the conventionally reared ewes, on which no lesions resembling orf were observed at any time during the study.

Historically, subtype B viruses in men who have sex with men (MSM) and injecting drug users (IDU) dominated the HIV epidemic in the United Kingdom, whereas non-B heterosexual infections dominate globally. Heterosexual contact is now the most common route of transmission in the United Kingdom. Here we monitor HIV subtype in Scotland, and link it to origin of infection. HIV-1 sequence was generated from new diagnoses and the subtype thus obtained linked with demographic data. Virus was subtyped from 80% (137/171) of all new diagnoses in Scotland. Of 58 individuals infected by heterosexual contact, 74% (43) harboured non-B viruses, contrasting with 7% (5/68) of those infected by IDU or MSM. Eighty-four per cent of non-Bs (46/55) were probably acquired outside the United Kingdom, but nine individuals probably acquired their non-B infection in the United Kingdom. Non-B subtypes of HIV-1 predominate in recently diagnosed, heterosexually acquired infections in Scotland and are present in all risk groups, even those with no exposure outside the United Kingdom.

Our objective was to monitor changes in the subtypes of HIV-1 infecting a rural Ugandan cohort where the spread of HIV-1 is by unprotected heterosexual contact and subtypes A and D predominate. Should one subtype be better able to spread we would anticipate a rise in incidence of one subtype at the expense of the other over a decade of study. We employed a natural history cohort, which had been established by the Medical Research Council in 1990 and subtyped virus from 90% (139) incident cases by DNA sequencing in two separate genes. We found that viral subtype had no predilection for males, females or age at infection and that between 1990 and 2000 there was no significant change in the relative number of different subtypes. The only significant trend was a reduction in the proportion of viruses classified as recombinant. This may reflect the overall decline in prevalence of HIV-1 in Uganda over this period.

International audience; Clinical studies show that in the absence of anti-retroviral therapy an immune response against the human immunodeficiency virus type 1 (HIV-1), transacting transcriptional activator (Tat) protein correlates with long term non-progression. The purpose of this study is to try to understand what can trigger an effective immune response against Tat. We used five Tat variants from HIV strains identified in different parts of the world and showed that mutations of as much as 38% exist without any change in activity. Rabbit sera were raised against Tat variants identified in rapid-progressor patients (Tat HXB2, a European variant and Tat Eli, an African variant) and a long term non-progressor patient (Tat Oyi, an inactive African variant). Enzyme-linked immunosorbent assay (ELISA) results showed that anti-Tat Oyi serum had the highest antibody titer and was the only one to have a broad antibody response against heterologous Tat variants. Surprisingly, Tat HXB2 was better recognized by anti-Tat Oyi serum compared with anti-Tat HXB2 serum. Western blots showed that non-homologous Tat variants were recognized by antibodies directed against conformational epitopes. This study suggests that the primary and tertiary structures of the Tat variant from the long term non-progressor patient are critical to the induction of a broad and effective antibody response against Tat.

Objective: To investigate the number and variety of viruses with discrepant subtypes between env and gag and within gag in two cohorts in Uganda. Methods: Sequences were generated from PCR products amplified directly (without cloning) from patient blood and compared in the v3/v4 region of env and the p17 and p24 regions of gag to reference subtype strains by phylogenetic analysis. Gag sequences with a discrepant subtype between p17 and p24 were analyzed further to indicate approximate sites of recombination. Results: Envelope subtypes D and A were predominant but subtypes B C and G were also found. From analysis of three short regions of the HIV genome we found 15 different combinations of subtype assortment including 11 different recombinant permutations. Approximately 30% of viruses (29/104) in this part of Uganda appear to be recombinants between the env and gag genes and 10% (11/104) are recombinant within the gag gene. There was no clear pattern of crossover points within the gag gene. There seems to be no evidence of new circulating recombinant forms. Conclusion: Both inter-genic and intra-genic inter-subtype recombination appear to be a relatively common occurrence in this geographical region where two subtypes of virus co-circulate. These results have implications for cross-clade vaccine design. (author’s)

OBJECTIVE: To investigate the role of HIV-1 envelope subtypes on disease progression in a rural cohort of Ugandan adults where two major HIV-1 subtypes (A and D) exist. METHODS: Participants of a clinical cohort seen between December 1995 and December 1998 had blood collected for HIV-1 subtyping. These included prevalent cases (people already infected with HIV at the start of the study in 1990) and incident cases (those who seroconverted between 1990 and December 1998). HIV-1 subtyping was carried out by heteroduplex mobility assay and DNA sequencing in the V3 env region. Disease progression was measured by the rate of CD4 lymphocyte count decline, clinical progression for the incident cases as time from seroconversion to AIDS or death, to first CD4 lymphocyte count < 200 x 10(6)/l and to the World Health Organization clinical stage 3. All analyses were adjusted for age and sex. RESULTS: One hundred and sixty-four individuals, including 47 prevalent and 117 incident cases, had V3 env subtype data of which 65 (40%) were subtyped as A and 99 as D. In the incident cases, 44 (38%) were subtyped as A and 73 as D. There was a suggestion that for most end-points A had a slower progression than D. The cumulative probability of remaining free from AIDS or death at 6 years post-seroconversion was 0.72 [95% confidence interval (CI), 0.50 to 0.85] for A and 0.58 (95% CI, 0.42 to 0.71) for D, and the adjusted hazard ratio of subtype D compared to A was estimated to be 1.39 (95% CI, 0.66 to 2.94; P = 0.39). The estimated difference in rates of decline in square root CD4 lymphocyte counts was -0.41 per year (95% CI, -0.98 to 0.15; P = 0.15). CONCLUSION: This study suggests that although subtype A may have a slower progression than D, HIV-1 envelope subtype is not a major factor in determining the progression of HIV-1 disease in a rural population in Uganda.

A pilot study was undertaken with the objective of developing a simple, economical, and efficient algorithm through which to subtype HIV-1 in a large epidemiological cohort study in Uganda. A peptide enzyme immunoassay (PEIA) employing both V3 and gp41 regions and a heteroduplex mobility assay (HMA) were evaluated in comparison with DNA sequencing. Of 146 samples selected, 115 (79%) were successfully sequenced. Taking sequence data as the "gold standard," other assays were compared with these data. The HMA correctly identified 95 (83%) of the samples, and only 1 sample was wrongly identified. The V3 PEIA alone and in combination with gp41 peptides correctly identified 76 and 78% of the samples, respectively; however, the number of wrongly identified samples was four times less with the combination compared with V3 peptides alone (4 versus 16%). The sensitivity, specificity, and positive and negative predictive values for serotype A and D samples were greater for the combination than V3 peptides alone. We have described a new algorithm to segregate subtypes A and D. This algorithm uses the two peptide assays followed by HMA and then DNA sequencing for untypable samples, giving an accuracy of 95% at a cost of 37 and 21% for consumables compared with subtyping all the samples by HMA or DNA sequencing, respectively. This proposed approach is suitable for epidemiological studies in Uganda and other regions with a predominance of A and D subtypes.

In a molecular investigation into the outbreak of HIV in Glenochil during the first 6 months of 1993, we previously demonstrated that 13 out of the 14 HIV positive inmates were infected with a virtually identical strain, and discounted 2 others as potential sources. Here we investigate a further 8 potential contacts and 4 potential sources which were identified in the companion paper [1]. We were able to examine viral sequence from all but one of these 12 and results have revealed them to be distinct both from each other and the original 14. Thus, despite an intensive follow-up investigation, we have been unable to identify any further HIV infections that might have been part of the 1993 outbreak. It is possible that persons who were infected at that time remain undetected; however this and the companion report strongly suggest that if this were the case the likely numbers would be few.

Summary Orf is a disease of sheep and goats which is caused by a parapox virus. It can be transmitted to human.s, and is considered an occupational hazard by those handling sheep. In this paper we present the first report of both cell-mediated and humoral immune responses to naturally acquired orf virus infection in humans. Lymphoproliferative responses of peripheral blood mononuclear cells of patients to an orf virus antigen were vigorous soon after infection, but rapidly declined. Orf virus antibody levels, detected by KLISA, were shown to rise during infection. Western blot analysis confirmed this, and demonstrated that the antibody produced in response to the infection was directed against the 40-kDa viral surface tubule protein. Where direct comparisons were possible, the immune response of humans to orf virus infection was similar to that previously reported for sheep. Evidence was obtained suggesting that prior exposure to vaccinia virus (smallpox vaccination) provided no protection from subsequent orf virus infection. In addition, orf virus infection did not enhance immune responses to vaccinia virus antigens.

Background: The SARA trial recently demonstrated a non-inferior CD4 response, over median follow-up of 60 weeks, with a boosted protease inhibitor monotherapy (bPImono) maintenance second-line regimen compared with continuous combination therapy (CT), suggesting this approach could maintain effectiveness whilst improving tolerability and decreasing costs [International AIDS Conference 2010, LBPE16]. Analysis of virological response and genotypic drug resistance is reported here. Methods: Eligible participants in the DART trial who received 24 weeks of lopinavir/ritonavir-containing second-line CT were randomised to maintain current CT or to reduce to bPImono within a nested pilot trial (SARA). No real-time virology was performed, but stored plasma samples from time at switch to second-line, randomisation after 24 weeks of second-line, and 24 weeks after randomisation were assayed for HIV-1 RNA viral load (VL) by Roche Amplicor v1.5. Genotypic resistance was assessed on samples with VL >1000 c/ml at this latest time point, along with paired samples at switch to second-line. All analyses are intention-to-treat. Results: 192 participants were randomised to CT (n=95) or bPImono (n=97). 77% (135/173) had VL 1000 c/ml at week 24, 12 (2 CT, 10 bPImono) have been assessed genotypically. IAS major PI mutations at week 24, not present at switch to second-line, were detected in 2 bPImono participants only. One participant (VL=3600 c/ml) had I54V only, the other (VL=1490 c/ml) M46IM+V82AV. Both isolates were considered fully susceptible to darunavir. Conclusions: In this study based on retrospective virological testing, bPImono following 24 week second-line induction was associated with an increase in low level viraemia, although generally in the absence of PI resistance. Longer-term trials are required before definitive conclusions can be drawn about the effectiveness of PI monotherapy in populations without access to virological monitoring. Supplement: Abstracts of the Tenth International Congress on Drug Therapy in HIV Infection http://www.biomedcentral.com/content/pdf/1758-2652-13-S4-info.pdf Conference: Tenth International Congress on Drug Therapy in HIV Infection 7-11 November 2010 Glasgow, UK (Published: 8 November 2010) doi:10.1186/1758-2652-13-S4-O20 Cite this article as: Pillay et al.: O214. Virological findings from the SARA trial: boosted PI monotherapy as maintenance second-line ART in Africa. Journal of the International AIDS Society 2010 13(Suppl 4):O20. Full text: PubMed Central: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3112833/

Functional and phenotypic changes in the cell populations were monitored in the popliteal efferent lymph of sheep following experimental epidermal infection with orf virus. In another group of sheep, cells from the popliteal lymph node draining the site of infection were similarly monitored and compared with the cells from contralateral popliteal and mesenteric lymph nodes. All sheep showed serological evidence of previous exposure to orf virus. Following infection, anti-orf antibody titres rose and efferent lymphocyte and blast cell output increased. Interferon-like activity was detected in efferent lymph early after orf virus but not mock infection. Lymphocytes from the draining popliteal lymph node showed antigen-specific lymphoproliferation on Days 3–7 while cells in the efferent lymph demonstrated proliferative activity on Days 4–6. The requirement for exogenous antigen-presenting cells in the culture of efferent lymphocytes varied between individual sheep. The culture supernatant from proliferating lymph node cells contained interferon-like activity but no anti-orf antibodies, the reverse of that from cultured efferent lymphocytes, perhaps indicating a different reactive T cell population. During the course of the experiment there was an increase in the percentage of efferent lymphocytes expressing MHC Class II antigens and surface immunoglobulins, the latter being recorded as a double peak. The short-term nature of the local T cell response may in part explain the incompleteness of immunity to orf virus in sheep.

Objectives: To support already established epidemiological links between inmates of Glenochil prison positive for HIV infection by using molecular techniques and thus provide evidence of the extent of acquisition during a recent outbreak of the disease resulting from needle sharing. To identify possible sources of the outbreak, and to demonstrate the ability of the methodology to make further links beyond the original outbreak. Design: Viral sequences obtained from the blood of HIV positive prisoners previously identified by standard epidemiological methods were compared with each other and with sequences from other Scottish patients. Setting: Glenochil prison for men, central Scotland. Subjects: Adult inmates and their possible contacts. Results: Phylogenetic analysis of viral sequences in two different genomic regions showed that 13 of the 14 HIV positive prisoners had been infected from a common source. Previous research had shown that six of these had acquired their infection in Glenochil; molecular evidence suggests that more than double this number were infected while incarcerated. Virus from two long term HIV positive patients who were in the prison at the time of the outbreak but who were not identified in the original or subsequent surveys was sufficiently different to make it unlikely that they were the source. A viral sequence from heterosexual transmission from one inmate showed the ability of these techniques to follow the infection through different routes of infection. Conclusion: The number of prisoners infected with HIV during the 1993 outbreak within Glenochil prison was more than twice that previously shown. This shows the potential for the spread of bloodborne diseases within prisons by injecting drugs. Key messages Originally, standard serological studies established that six out of 14 HIV positive people had acquired their infection in Glenochil prison in 1993 A subsequent survey estimated that up to 20 prisoners were HIV positive at that time Molecular linkage techniques showed that 13 of the original 14 infected men had viral sequences similar enough to indicate that one source of infection was common to all Molecular techniques provide a powerful epidemiological tool either to link or to distance infections with HIV Injecting drug use is a potentially explosive health care problem in prisons

To assess the effect of mutations at the CCR-2 and CCR-5 loci on heterosexual human immunodeficiency virus (HIV) transmission, 144 persons heterosexually exposed to HIV (infected and uninfected [EU]) and 57 HIV-positive index partners were genotyped. A significantly higher frequency of 64I heterozygotes at CCR-2 was observed in HIV-positive than in EU women (P=.02, relative risk=1.6). The allele frequency of 64I in women was 8% in HIV-positive contacts and 1% in EUs (P