Characterization of CD8+ T-cell response in acute and resolved hepatitis A virus infection.

BACKGROUND & AIMS: In contrast to the infection with other hepatotropic viruses, hepatitis A virus (HAV) always causes acute self-limited hepatitis, although the role for virus-specific CD8 T cells in viral containment is unclear. Herein, we analyzed the T cell response in patients with acute hepatitis by utilizing a set of overlapping peptides and predicted HLA-A2 binders from the polyprotein. METHODS: A set of 11 predicted peptides from the HAV polyprotein, identified as potential binders, were synthesized. Peripheral blood mononuclear cells (PBMCs) from patients were tested for IFNγ secretion after stimulation with these peptides and ex vivo with HLA-A2 tetramers. Phenotyping was carried out by staining with the activation marker CD38 and the memory marker CD127. RESULTS: Eight out of 11 predicted HLA-A2 binders showed a high binding affinity and five of them were recognized by CD8+ T cells from patients with hepatitis A. There were significant differences in the magnitude of the responses to these five peptides. One was reproducibly immunodominant and the only one detectable ex vivo by tetramer staining of CD8+ T cells. These cells have an activated phenotype (CD38hi CD127lo) during acute infection. Three additional epitopes were identified in HLA-A2 negative patients, most likely representing epitopes restricted by other HLA-class I-alleles (HLA-A11, B35, B40). CONCLUSIONS: Patients with acute hepatitis A have a strong multi-specific T cell response detected by ICS. With the tetramer carrying the dominant HLA-A2 epitope, HAV-specific and activated CD8+ T cells could be detected ex vivo. This first description of the HAV specific CTL-epitopes will allow future studies on strength, breadth, and kinetics of the T-cell response in hepatitis A.

The epidemiology of oro-nasal haemorrhage and suffocation in infants admitted to hospital in Scotland over 10 years.

OBJECTIVE: To estimate the incidence of oro-nasal haemorrhage (ONH) and suffocation in infancy and to investigate their aetiology and overlap. Setting A 10-year retrospective hospital based study from Scotland, UK. METHODS: The hospital notes of all infants presenting with ONH or suffocation identified through the Information Services Division of the Scottish Health Service were reviewed by three paediatric consultants, two with child protection expertise. The hospital-based incidences of haemorrhages from different sites were calculated and the causes ascertained. When trauma was involved, a decision was made whether it was likely to have been accidental. RESULTS: 7 cases of suffocation and 88 of ONH were recorded at hospital discharge over 10 years. This gives an incidence of ONH of 1.62 (1.30 to 1.99)/10,000 live births (95% CI) which consists of haemorrhage arising from nose or mouth (N/M), n=65 (1.19/10,000 (0.92 to 1.52)); haematemesis, n=11 (0.20/10,000 (0.10 to 0.36)); haemoptysis, n=3 (0.06/10,000 (0.1 to 0.16)) and pulmonary haemorrhage, n=9 (0.17/10,000 (0.08 to 0.31)). No suffocation cases were recorded as having a coincident ONH, but five ONH cases were probably caused by airway obstruction. 40 of 65 cases of N/M were associated with trauma, which in 15 cases were thought to be probable abuse; four were associated with coagulation abnormalities. 2/3 haemoptysis cases, 2/11 haematemesis cases and 8/65 N/M cases were associated with a coincident respiratory tract infection, though in 4/8 of these cases, there was an associated apparent life-threatening event. CONCLUSIONS: Haemorrhage from the N/M is rare in infancy. Trauma is commonly involved and child protection concerns often poorly explored. Pulmonary haemorrhage and several cases of ONH were associated with probable airway obstruction. Information, in cases of ONH, is in general recorded badly, and an investigation and management plan are suggested.

Non-accidental head injury: a consequence of deprivation?

BACKGROUND: Non-accidental head injury (NAHI) is a significant personal and public health problem, with considerable mortality and morbidity. The evidence base for risk factors specific for NAHI is limited due to difficulties with case definition and study design. The risk factors associated with NAHI in infants was evaluated in this study, and the extent to which indices of deprivation influence this health problem was addressed. METHODS: A 10-year prospective study was conducted in Scotland involving all paediatric hospitals and other general hospital departments admitting children. Subjects were children ≤2 years of age, with a diagnosis of "suspected NAHI". Socioeconomic characteristics of the index cases were compared to the general population, using the Scottish Index of Multiple Deprivation (SIMD) 2006. RESULTS: There were highly significant differences (p<0.001) between the SIMD rank scores of the NAHI cases and scores for the whole Scottish population. For the cohort, SIMD ranks ranged from 34 to 6253 (median 1210; mean 1577) compared to the population range of 1-6505 (median and mean=3253). Similar differences were found for each of the component domains of income, employment, health, education, crime and housing (p<0.001). In contrast, the scores for "geographic access" (to essential service) were higher than for the whole population (p<0.001), indicating that the deprivation was not due to lack of local services. CONCLUSION: In Scotland, children who present with suspected NAHI originate predominantly from the most deprived areas of the community. Public health and intervention strategies should be focused in these areas.

The dental practitioner and child protection in Scotland.

OBJECTIVES: To identify from general dental practitioners: undergraduate and postgraduate training experience in child protection; numbers of suspected cases of child physical abuse; reasons for failing to report suspicious cases of child physical abuse; knowledge of local child protection protocols and procedures for referral. MATERIALS AND METHODS: Postal questionnaires were sent to 500 randomly selected general dental practitioners in Scotland, with a further 200 sent to a random sample of the original 500 to increase response rate. RESULTS: Sixty-one per cent (306) of the original 500 questionnaires, and 35% (69) of the second random mail shot of 200 questionnaires were returned. Only 19% could remember any undergraduate training and 16% had been to a postgraduate lecture or seminar in child protection. Twenty-nine per cent of dentists had seen at least one suspicious case in their career. Only 8% of suspicious cases were referred on to the appropriate authorities. Reasons for failure to refer revealed that 11% were concerned about a negative impact on their practice, 34% feared family violence towards the child, 31% feared violence directed against them, and 48% feared litigation. Only 10% of dentists had been sent a copy of the local child protection guidelines on commencing work and only 15% had seen their Area Child Protection Committee (ACPC) Guidelines via any route. CONCLUSIONS: Due to lack of training or clear guidelines for dentists in Scotland, most practitioners were unsure what to do in the event of a suspicion of child abuse. Twenty-one per cent of dentists had encountered suspicious cases but failed to take any action. Dentists overwhelmingly requested appropriate training. This training should address dental competence in assessment of suspicious indicators and involve dentists in inter-agency child protection training.

Injuries to the head, face, mouth and neck in physically abused children in a community setting.

OBJECTIVES: The aims of the present study were to identify the incidence of orofacial injuries found within a cohort of physically abused children, and examine demographic data surrounding the alleged perpetrator, the location in which the alleged assault occurred, the mechanism of injury and the actual orofacial injury incurred. METHODS: The research took the form of a retrospective study of clinical case records of children with suspected physical abuse from 1 June 1998 to 31 May 2003. Seven hundred and fifty case records were identified and 390 (46.7%) were available for data extraction. RESULTS: Fifty-nine per cent (n = 230) of children had signs of abuse on the head, face or neck. The alleged perpetrator was the mother in 104 cases (26.7%), the father in 100 (25.6%) and mother's partner in 49 other cases (12.6%). More than half (53.3%) of the alleged abuse occurred in the child's home; in 32.3% of cases, the location was not recorded. Other locations included outside in a public place, school and at the home of the alleged abuser. Some 23.4% (n = 54) had been punched or slapped around the head, neck or face, 17.4% (n = 40) had been struck by an object, and 15.2% (n = 35) had allegedly sustained multiple modes of injury. Bruising to the head, neck or face was seen in 95.2% (n = 219) of children, and 32.6% (n = 75) had abrasions; 65.2% (n = 150) of the bruises and 22.9% (n = 53) of the abrasions were on the face. CONCLUSIONS: Fifty-nine per cent of physically abused children in the present cohort had orofacial signs of abuse which would be easily visible to a dental practitioner. The commonest injuries were bruises and abrasions. This concurs with previous reports in the literature and highlights the important role of dental practitioners in the recognition of children who have been abused.

Lack of agreement on colour description between clinicians examining childhood bruising.

INTRODUCTION: In child protection cases clinicians are often asked to describe and age bruises. This paper looks at both intra- and inter-observer variability in the description of childhood bruising. METHODS: Fifty-eight bruises on 44 children were described by three observers, the bruises were then photographed and the same observers described the bruises at a later date. The descriptions were compared and classified in terms of complete, partial, or no agreement, both between observers and between the in vivo and photographic descriptions. RESULTS: Complete agreement on colour description between two observers in vivo occurred in 27% of descriptions in vivo and 24% of photographs. Only 31% of descriptions completely agreed with the later description of a photograph of the same bruise. CONCLUSIONS: This marked variability in colour description, severely questions the practice of estimating the age of bruises especially from clinical photographs as evidence in child protection proceedings.

In vitro measurement of cytotoxic T cell activity does not predict clinical progression in paediatric HIV disease--two case studies.

Cytotoxic T cells are believed to be an important immune response in HIV infection, both in the initial response to viraemia, and in controlling HIV replication and maintaining clinical stability. We report here the detailed findings in two vertically infected children, from the Edinburgh perinatal cohort. Both were clinically stable for the first 7 years of life. One had vigorous HIV-specific cytotoxic T lymphocyte (CTL) responses, and non-lytic suppression, measured in vitro, while the second had no CTL activity against HIV. Despite her HIV-specific immunity, the first child had a declining CD4 count, and a high and fluctuating viral load, whereas the second child maintained a stable CD4 count, a low viral load and had a virus which could not be cultured in peripheral blood mononuclear cells (PBMC) in vitro. The first child subsequently progressed to AIDS and has now died, while the second remains clinically well. More detailed investigations showed the clinically stable child to be heterozygous for the CCR5 receptor, and to be HLA-B49--both of which markers have been associated with slow HIV disease progression. These findings question the role of CTL in maintaining stable HIV disease, and stress the need for immunological investigations to be considered in the light of the genetic make-up of the patient. They may also reflect a different immunopathogenesis of HIV disease in children compared with adults.

Maternal HIV and drug use: effect on health and social morbidity.

A retrospective analysis of routine child health surveillance information was performed on health visitor records of 459 children, to examine the independent effects of maternal HIV infection and drug use during pregnancy on morbidity in the first 3 years of life. No significant differences were observed in the developmental progress of children born to HIV infected or drug using women when compared to community controls. The pattern of medical consultations in the first 18 months of life was significantly different, maternal drug use exerting a negative influence on outpatient visits (odds ratio 0.6, 95% confidence interval 0.4 to 1.0). At 6 weeks, the majority of children lived with their birth parent(s), and no differences were observed between the groups. By 10 months of age, only 81% of children born to HIV infected drug using women lived with their parent(s). While maternal drug use and HIV did not have adverse effects on child health and development, these findings highlight the social implications for children affected by the heterosexual spread of HIV.

CD4 and CD8 lymphocytes in diagnosis and disease progression of pediatric HIV infection.

Vertical infection with human immunodeficiency virus-1 (HIV-1) causes profound changes in the proportions of subpopulations of lymphocytes in the peripheral circulation. In this study the percentages in whole blood of CD4 and CD8 cells, and of immunologically important subpopulations, were measured in 19 HIV-infected children over periods of up to 4 years and compared to our recently published ranges for normal children of various ages. The rate of CD4 decline and of CD8 increase differed between clinically fast and slow progressors. On CD8 cells, cytotoxic, memory (CD11abright and CD45R0), and activation (HLA-DR) markers were raised soon after birth to levels outside the normal range, and compared favorably with HIV culture as a method for early diagnosis of HIV infection. Mean levels of naive (CD45RA) and memory (CD45R0, CD29) markers on CD4 cells became significantly altered after 48 months of age, suggesting that these are markers of more advanced disease. Despite different ages of enrollment into the study, in the cohort as a whole, the levels of the lymphocyte subpopulations studied changed consistently. Thus, their measurement could be useful both in the diagnosis and prognosis of HIV infection in individual children. This is the first report showing that lymphocyte subpopulation analysis can play a major role in the diagnosis of pediatric HIV infection.

Age-related ranges of memory, activation, and cytotoxic markers on CD4 and CD8 cells in children.

The expression of markers defining functional subpopulations on the surface of CD4 and CD8 cells changes with disease. To monitor these changes in children, it is important to establish the age-related normal changes in marker expression due to maturation of the immune system. We have studied the expression of several functionally important molecules on both CD4 and CD8 cells in 168 children (aged 0-122 months) using monoclonal antibodies and flow cytometry. Our results show that the percentage of CD4 cells decreases with age, while the CD8 percentage increases, resulting in a decrease in the CD4/CD8 ratio. The expression of CD45RO and CD29 increases with age, while CD45RA expression decreases, both on CD4 and CD8 cells. The expression of HLA-DR on both CD4 and CD8 cells, and of CD11a and CD57 on CD8 cells, is less clearly age dependent. The relationships between the marker percentages and age were not straightforward; the standard deviations and the skewness, as well as their mean values, varied as a function of age. The changes were modeled for each marker and age-specific centiles are presented.

Cytotoxic T lymphocyte activity and CD8 subpopulations in children at risk of HIV infection.

HIV-specific cytotoxic T lymphocytes (CTL) are thought to play a major role in viral control in HIV-infected adults. Changes in the relative proportions of CD8 lymphocyte subpopulations are also thought to be associated with disease progression. Less is known about the relative effectiveness of CTL against different HIV targets, or about the relationship, if any, between CTL activity and CD8 subpopulations. We have measured CTL activity against four HIV gene products (gag, tat, pol and env) and expression of CD45RO, CD45RA, HLA-DR, CD29, S6F1, and CD57 surface markers on CD8 cells from nine HIV-infected and 11 HIV-uninfected children. Of nine HIV-infected children, six showed antigen-specific CTL activity on at least one occasion: 4/6 directed against tat, 6/6 against pol, 1/6 against env, and 1/6 against gag. However, the specificity of the CTL activity varied between children and within individual children with time. Furthermore, two uninfected children showed CTL activity, one to HIV-gag, -pol and -tat, and the other to HIV-pol. All the HIV-infected and two uninfected children had abnormal proportions of CD8 subpopulations in whole blood compared with age-matched controls. There was no correlation between CTL activity and CD8 subsets in whole blood. Five children changed from CTL-positive to CTL-negative (or vice versa) during the study. In these, the occasions when CTL activity was detected coincided with an increase in CD8 cells, an expansion of HLA-DR+ CD8 cells and a loss of CD45RA+ CD8 cells.

HIV testing and prevalence in pregnancy in Edinburgh.

The objective was to study the changes in pregnancy HIV prevalence with time. Data were collected from multiple sources to provide a comprehensive record of all HIV seropositive pregnant women identified in the Edinburgh area (Scotland) until December 1992. There were 177 pregnancies in 108 HIV seropositive identified women. Risk factors were injection drug use (79% of pregnancies) and a known HIV seropositive injection drug-using partner (16%). Prevalence has decreased for Edinburgh City women from 0.5% of all pregnancies in 1986 to 0.1% in 1992; It was higher for induced abortion (0.6%) than for delivery (0.2%). HIV testing in pregnancy has declined. Comparison with unlinked anonymized testing showed that in 1990-1991, 20/22 seropositive women were known. In 1992, only 3 of 10 seropositive pregnancies were identified. The cohort initially infected by exposure to a 'drug related' risk factor between 1983 and 1985 may have increasingly finished childbearing, deliberately decided against pregnancy because of HIV status, and declined because of death, illness and emigration from the area, There may not have been major early tertiary heterosexual spread; however, data from 1992 suggest that this could now be impacting on pregnancy prevalence. Local testing policies have not adapted to this possible change.

Cytotoxic T lymphocyte activity in children infected with HIV.

Of the Edinburgh cohort of approximately 130 children born to HIV-infected women, 9 are infected and alive. This article describes results from the first 18 months of a natural history study of seven of these, and two adopted children, studying the CD8 T cell-mediated cytotoxicity against HIV proteins (Gag, Tat, Pol, and Env), over time, and relating it to clinical progression and viral activity. Autologous EBV cell lines infected with vaccinia-HIV constructs were used as target cells, and bulk-cultured peripheral blood mononuclear cells as effector cells. The children ranged in age from 0 to 93 months, with six of the nine showing CTL activity to one or more HIV proteins. The specificity of the response was directed against Tat in the younger children, switching to Pol, then Gag or Env. Preliminary analysis of virological data showed no association between CTL and virus activity. The children with CTLs tended to be well clinically, but the cohort needs to be studied longer before conclusions can be made about CTL activity and HIV disease progression. Cytotoxic T lymphocyte activity has also been observed in two children diagnosed as HIV uninfected. These results show the importance of looking at CTL specificity, and may have implications in vaccine design.

Measles, mumps and rubella immunisation in children at risk of infection with human immunodeficiency virus.

A retrospective serological study of 81 children at risk of HIV (II HIV-infected, 70 HIV-negative) was performed to assess susceptibility to measles, mumps and rubella and response to measles, mumps and rubella (MMR) immunisation. There was no difference in the initial serological response between the HIV-infected and HIV-negative children. Repeat serology should be performed on HIV-infected children as during follow-up antibodies may be lost. Reimmunisation should be considered for the seronegative.

Maternal factors in HIV transmission.

In order to identify features associated with an increased risk of transmission of HIV from seropositive women to their offspring, 70 children of 58 HIV seropositive mothers were studied. Fifty-six children were followed prospectively from pregnancy; in 14 identified after the puerperium, obstetric notes were reviewed and stored serum was tested. Twelve infants of 10 mothers were HIV infected. Risk of transmission was increased in the first year after seroconversion; 5/9 infants born at this time were infected compared with 7/61 born subsequently (P < 0.001). Progression to stage IV in transmitters was more likely, occurring in the mothers of 9 infected children at a median of 3 years (range 0.5-6.5) and in mothers of 19 non-infected children at a median of 5 years (range 1-7) (P = 0.032). Maternal CD4+ counts < 400 x 10(6)/l were found in 7/12 transmitting and 7/49 non-transmitting pregnancies (P < 0.01). Differences in HIV antigenaemia did not reach significance. These factors may influence the counselling of mothers regarding their child's and their own prognosis.

Dizygotic twins discordant for HIV and hepatitis C virus.

Twin girls were born at 37 weeks' gestation to a mother infected by HIV and hepatitis C virus. Twin 1 had symptomatic HIV infection by 9 months but was negative for hepatitis C virus antibody and RNA. Twin 2 became HIV antibody negative by 15 months but was positive for antihepatitis C virus and RNA.

Infrequent vertical transmission of hepatitis C virus.

The rate of vertical transmission of hepatitis C virus (HCV) was determined by a combination of assays for anti-HCV antibody and the polymerase chain reaction (PCR) in 66 children born to infected mothers. Only 4 children showed evidence of infection with HCV, being positive for anti-HCV in all samples collected from 6 months to 5 years of age. All samples from the remaining 62 children were repeatedly anti-HCV-negative on screening by two second-generation antibody assays. Furthermore, samples collected at age 12 months from 30 antibody-negative children born of HCV-infected mothers were uniformly PCR-negative, showing that "seronegative" infection with HCV was rare or absent in this study group. Serologic reactivity to HCV-encoded antigens in samples from infected children was largely confined to the HCV core protein. Infection with human immunodeficiency virus in the mother was not a significant cofactor for mother-to-child transmission of HCV.

Cardiac dysfunction in paediatric HIV infection.

A child under long term surveillance for vertically acquired HIV-I infection developed respiratory symptoms. Dilated cardiomyopathy was diagnosed. Clinical and echocardiographic assessment of six other children with symptomatic HIV disease showed no evidence of cardiac dysfunction. The exact prevalence of this complication in paediatric HIV infection is unknown, but the potential diagnostic difficulties are highlighted.