The incidence of asthma and atopic dermatitis (AD) were evaluated in HIV-infected (n=451) compared to HIV-exposed (n=227) but uninfected (HEU) children and adolescents by abstraction from clinical charts. Asthma was more common in HIV-infected compared to HEU children by clinical diagnosis (25% vs. 20%, p = 0.101), by asthma medication use, (31% vs. 22%, p = 0.012), and by clinical diagnosis or both medication use, (34% vs. 25%, p = 0.012). HIV-infected children had a greater risk of asthma compared to HEU children (HR = 1.37, 95% CI: 1.01 to 1.86). AD was more common in HIV-infected than HEU children (20% vs. 12%, p = 0.009)) and children with AD were more likely to have asthma in both cohorts (41% vs. 29%, p = 0.010). HIV-infected children and adolescents in this study had a 30% increased incidence of asthma and AD, a finding critical for millions of HIV-infected children worldwide.
Atopic dermatitis; asthma; pediatric HIV infection; inhaled corticosteroids; protease inhibitors
Investigations of basic immunological mechanisms and clinical studies of primary immunodeficiencies were most prevalent in 2011. Significant progress was achieved in the characterization of TH17 cells differentiation and their associated cytokines in the setting of inflammatory disorders, HIV infection and immunodysregulation disorders. The role of TACI mutations in the pathogenesis of CVID was further described and reported to be likely mediated by impaired TACI expression affecting B cell function. The frequency of autoimmunity in partial DiGeorge syndrome was estimated at 8.5%, predominantly resulting in blood cytopenias and hypothyroidism. Several reports emphasized the presentation of neoplasias, most often lymphomas, as the first manifestation of several primary immunodeficiencies. Novel strategies for newborn screening of B cell lymphopenia by measuring immunoglobulin K chain deletion recombinant excision circles (KRECs) and for adenosine deaminase deficiency using tandem mass spectrometry were demonstrated to be feasible at large scale. Progress in the treatment of primary immunodeficiencies included increased success using unrelated HLA-compatible donors for hematopoietic stem cell transplantation, and the development of new gene therapy approaches providing with increased safety features. Induced pluripotent stem cells were developed from patients with primary immunodeficiencies, providing with a virtually unlimited resource for pathophysiology and gene correction studies. New findings in several of the uncommon immunodeficiencies, such as the increased susceptibility to severe viral infections secondary to defects in the activation of the toll-like receptor 3 pathway, overall contributed to the understanding of their immunological basis and provided for the design of effective diagnostic and therapeutic strategies.
Immunology; primary immunodeficiencies; IVIG; TACI; CVID; cell immunity; SCID; newborn screening
Goldenhar syndrome (GS) results from an aberrant development of the 1st and 2nd branchial arches. There is a wide range of clinical manifestations, the most common being microtia, hemifacial microsomia, epibulbar dermoids and vertebral malformations. We present two cases of GS and secondary immunodeficiency due to anatomical defects characteristic of this disorder. Case 1 (3-year-old female) averaged 6 episodes of sinusitis and otitis media per year. Case 2 (7-year-old female) also had recurrent otitis media, an episode of bacterial pneumonia, and 2 episodes of bacterial meningitis. Their immune evaluation included a complete blood count with differential, serum immunoglobulin levels and specific antibody concentrations, lymphocyte phenotyping, and mitogen and antigen responses, the results of which were all within normal ranges. Both children demonstrated major structural abnormalities of the inner and middle ear structures, retention of fluid in mastoid air cells, and chronic sinusitis by computed tomography. These two cases illustrate how a genetically-associated deviation of the middle ear cleft can cause recurrent infections and chronic inflammation of the middle ear and adjacent sinuses, even meninges, leading to a greatly reduced quality of life for the child and parents.
Goldenhar syndrome; Hemifacial microsomia; Oculo-auriculo-vertebral dysplasia; Recurrent infections; Sinusitis; Otitis; Meningitis; Immunodeficiency
Allogeneic hematopoietic cell transplantation (HCT) has been employed for 40 years to ameliorate or cure primary immune deficiency (PID) diseases, including severe combined immune deficiency (SCID) and non-SCID PID. There is a critical need for evaluation of the North American experience of different HCT approaches for these diseases, in order to identify best practices and plan future investigative clinical trials. A conference of experts in HCT treatment of PID has recommended: (1) a comprehensive cross-sectional and retrospective analysis of HCT survivors with SCID; (2) a prospective study of SCID patients receiving HCT, with comparable baseline and follow-up testing across participating centers; (3) a pilot study of newborn screening for SCID to identify affected infants prior to compromise by infection; and (4) for the non-SCID diseases, Wiskott-Aldrich syndrome and Chronic Granulomatous Disease, studies of the natural history of disease in patients who do or do not receive HCT. To accomplish these goals, collaboration by a consortium of institutions in North America is proposed. Participation of immunologists and HCT physicians having interest in PID and experts in laboratory methods, clinical outcomes assessment, databases and analysis will be required for the success of these studies.
Allogeneic hematopoietic cell transplantation; primary immunodeficiency; clinical trial
The effect of pre-transplant conditioning upon the long-term outcomes of patients receiving hematopoietic stem cell transplantation (HSCT) for severe combined immunodeficiency (SCID) has not been completely determined.
To assess the outcomes of 23 mostly conditioned SCID patients and compare their outcomes to 25 nonconditioned SCID transplanted patients previously reported.
In the present study, we reviewed the medical records of these 23 consecutive mostly conditioned SCID patients transplanted between 1998 and 2007.
Eighteen patients (median age at transplant 10 [range 0.8–108] mo) received haploidentical mismatched related donor, matched unrelated donor, or mismatched unrelated donor transplants, 17 of whom received pretransplant conditioning and one was not conditioned, 13 (72%) engrafted and survive a median of 3.8 [1.8–9.8] yr, 5 of 13 (38%) require IVIG, and 6 of 6 age-eligible children attend school. Of five recipients (median age at transplant 7 [range 2–23] mo) of matched related donor transplants, all 5 engrafted and survive a median of 7.5 [range 1.5–9.5] yr, 1 requires IVIG, and 3 of 3 age-eligible children attend school. Gene mutations were known in 16 cases: IL2γR in 7 patients, IL7αR in 4 patients, RAG1 in 2 patients, ADA in 2 patients, and AK2 in 1 patient. Early outcomes and quality of life of the previous non-conditioned vs. the present conditioned cohorts were not statistically different but longer-term follow-up is necessary for confirmation.
HSCT in SCID patients results in engraftment, long-term survival, and a good quality of life for the majority of patients with or without pre-transplant conditioning.
Hematopoietic stem cell transplantation; graft-versus-host disease; severe combined immunodeficiency; immune reconstitution; primary immunodeficiency; conditioning
Reports in basic and clinical immunology in 2010 reflected the use of state of the art genetic and immunological tools to characterize the pathogenesis of immunological diseases, and the development of novel therapies directed to these conditions. B cell biology has been explained in greater detail, significantly with lessons from the genetic defects found in the humoral immunodeficiencies. Monoclonal therapeutic antibodies are given for an increasing number of indications, such as anti-CD20 antibodies or Rituximab, which was initially developed for non-Hodgkin lymphomas and is currently used in diverse autoimmune and inflammatory disorders. The report of an infant with severe combined immunodeficiency in Massachusetts detected by newborn screening and successfully treated with hematopoietic stem cell transplantation validated recent efforts towards newborn screening for severe combined immunodeficiencies (SCID). Improvement of survival outcomes of primary immunodeficiencies patients treated with hematopoietic stem cell transplantation was demonstrated in a large European cohort with significant appreciation of the type of donor graft, particularly the use of HLA-matched unrelated donors for non-SCID patients. Progress in cellular mechanism of drug hypersensitivity included the characterization of nitroso-modified drug metabolites as potent T cell activators and the identification of the relocation of plasmacytoid dendritic cells from blood to skin as a potential risk factor for reactivation of viral disease.
Immunology; primary immunodeficiencies; B cell; Innate immunity; hereditary angioedema; drug allergy; cell immunity; SCID; newborn screening
To investigate the possible effects of antiretroviral therapy (ART) in utero on cardiac development and function in HIV-negative children.
ART reduces vertical HIV transmission. Long-term cardiotoxicity after in utero exposure to ART is unknown in children but has occurred in young animals.
Using a prospective multi-site cohort study design, we compared echocardiograms taken between birth and 24 months in two groups of HIV-negative infants of HIV-positive mothers: 136 infants exposed to ART (ART+) and 216 unexposed infants (ART−).
Mean LV mass Z-scores were consistently lower in ART+ girls than in ART− girls: differences in mean Z-scores were −0.46 at birth (P=0.005), −1.02 at 6 months (P<0.001), −0.74 at 12 months (P<0.001), and −0.79 at 24 months (P<0.001). Corresponding differences in Z-scores for boys were smaller: 0.13 at 1 month (P=0.42), −0.44 at 6 months (P=0.01), −0.15 at 12 months (P=0.37), and −0.21 at 24 months (P=0.21). Septal wall thickness and LV dimension were smaller than expected in ART+ infants, but LV contractility was consistently about 1 SD higher at all ages (P<0.001). In ART+ infants, LV fractional shortening was higher than in ART− infants; girls showed a greater difference.
Fetal exposure to ART is associated with reduced LV mass, LV dimension, and septal wall thickness Z-scores and increased LV fractional shortening and contractility up to age 2 years. These effects are more pronounced in girls than in boys. Fetal ART exposure may impair myocardial growth while improving depressed LV function.
Pediatric; HIV; Antiretroviral Therapy; Cardiomyopathy
Immunoreconstitution of HIV-infected (HIV+) patients after treatment with highly antiretroviral therapy (HAART) appears to provoke inflammatory diseases.
Determine whether HIV+ children on HAART (HIV+ HAART+) have a higher incidence of asthma than HIV+ children not on HAART (HIV+ HAART−).
To investigate this possibility, 2,664 children (193 HIV+, 2,471 HIV−) born to HIV+ women were evaluated for the incidence and prevalence of asthma (i.e., asthma medication use), and change of CD4+ T cell percentage with time.
The HIV+ HAART+ children had higher CD4+ T cell percentages, lower CD8+ T cell percentages, and lower viral burdens than the HIV+ HAART− children (P≤0.05 to P≤0.01). The cumulative incidence of asthma medication use in HIV+ HAART+ children at 13.5 year rose to 33.5% vs. 11.5% in HIV+ HAART− children (hazard ratio=3.34, P=0.01) and was equal to that in the HIV− children. In children born prior to the HAART era, the prevalence of asthma medication use for HIV+ HAART+ children at 11 years of age was 10.4% vs. 3.8% for HIV+ HAART− children (odds ratio=3.38, P=0.02) and was equal to that of the HIV− children. The rate of change of CD4+ T cells (percent/year) around the time of first asthma medication for HIV+ HAART+ vs. HIV+ HAART− children was 0.81 vs. −1.43 (P=0.01).
The increased incidence of asthma in HIV+ HAART+ children may be driven by immunoreconstitution of CD4+ T cells.
This HIV model of pediatric asthma may yield clues to help explain the epidemic of asthma in the general pediatric population.
pediatric HIV infection; CD4+ T cell mediated induction of asthma; HAART-produced immunoreconstitution
There are limited data on the immune profiles of HIV-positive children, compared with healthy controls, and no such data for Asian children.
To immunophenotype HIV-positive Asian children, including long-term non-progressors (LTNPs), compared with age-matched healthy controls.
We used flow cytometry to analyze 13 lymphocyte and monocyte subsets from 222 untreated, HIV-positive children with 15%–24% CD4+ T cells and no AIDS-related illnesses and 142 healthy children (controls). Data were compared among age categories. Profiles from LTNPs (n=50), defined as children≥ 8 years old with CD4+ T-cell counts ≥ 350 cells/mm3, were compared with data from age-matched non-LTNPs (n=17) and controls (n=53).
Compared with controls, HIV-positive children had lower values (cell count per mm3 and percent distribution) for helper T cells and higher values for cytotoxic T cells, with reductions in populations of naïve helper and cytotoxic T cells, B cells, and natural killer (NK) cells. HIV-positive children had high values for activated helper and cytotoxic T cells. Compared with non-LTNPs, LTNPs had higher values of helper and cytotoxic T cells, naïve and memory T-cell subsets, and B and NK cells. Surprisingly, counts of activated helper and cytotoxic T cells were also higher among LTNPs. LNTPs were more frequently male.
Untreated, HIV-infected Asian children have immune profiles that differ from those of controls, characterized by low values for helper T cells, naive T cells, B cells, and NK cells but high values for cytotoxic, activated helper, and cytotoxic T cells. The higher values for activated T cells observed in LTNPs require confirmation in longitudinal studies.
The distinct immunologic profile of LTNPs might identify lymphocyte subsets associated with HIV disease progression.
HIV; children; lymphocyte; monocyte; phenotyping; long-term non-progressors; antiretroviral therapy; Asia; disease progression; pediatric AIDS
The rapid response system (RRS) is a process of accessing help for health professionals when a patient under their care becomes severely ill. Recent studies and meta-analyses show a reduction in cardiac arrests by a one-third in hospitals that have introduced a rapid response team, although the effect on overall hospital mortality is less clear. It has been suggested that the difficulty in establishing the benefit of the RRS has been due to implementation difficulties and a reluctance of clinical staff to call for additional help. This assertion is supported by the observation that patients continue to have poor outcomes in our institution despite an established RRS being available. In many of these cases, the patient is often unstable for many hours or days without help being sought. These poor outcomes are often discovered in an ad hoc fashion, and the real numbers of patients who may benefit from the RRS is currently unknown. This study has been designed to answer three key questions to improve the RRS: estimate the scope of the problem in terms of numbers of patients requiring activation of the RRS; determine cognitive and socio-cultural barriers to calling the Rapid Response Team; and design and implement solutions to address the effectiveness of the RRS.
The extent of the problem will be addressed by establishing the incidence of patients who meet abnormal physiological criteria, as determined from a point prevalence investigation conducted across four hospitals. Follow-up review will determine if these patients subsequently require intensive care unit or critical care intervention. This study will be grounded in both cognitive and socio-cultural theoretical frameworks. The cognitive model of situation awareness will be used to determine psychological barriers to RRS activation, and socio-cultural models of interprofessional practice will be triangulated to inform further investigation. A multi-modal approach will be taken using reviews of clinical notes, structured interviews, and focus groups. Interventions will be designed using a human factors analysis approach. Ongoing surveillance of adverse outcomes and surveys of the safety climate in the clinical areas piloting the interventions will occur before and after implementation.
In 2009, reports on basic and clinical immunology had an increased focus on human disease mechanisms and management. The molecular pathogenesis of familial angioedema associated with estrogen was further explored to find possible factors affecting severity, including polymorphisms in enzymes and receptors related to bradykinin pathways. A placebo-controlled clinical trial of C1INH concentrate in hereditary angioedema patients demonstrated the safety of its use and the efficacy to reduce duration of angioedema attacks. The interaction of innate immunity and adaptive responses was further examined in several reports establishing the significant role of toll-like receptor stimulation for the development of optimal specific antibody responses. The 2009 update of the classification of primary immunodeficiencies introduced over 15 new genetic defects related to the immune response, including DOCK8 mutations responsible for the autosomal recessive form of the hyper IgE syndrome. Other reports expanded the clinical spectrum of disease and improved the characterization of conditions such as warts, hypogammaglobulinemia and myelokathexis (WHIM) syndrome, or the occurrence of mucormycosis and Serratia infections in chronic granulomatous disease. The frequent presentation of gastrointestinal disorders in humoral immunodeficiencies was recognized and recommendations for management were reviewed. Clinical research focused in severe combined immunodeficiency included the development and implementation of a state-wide newborn screening program for this condition, a desired goal considering the significant reduction of mortality rate when diagnosis is made early before opportunistic infections occur.
basic immunology; B cells; T regulatory cells; TH17; clinical immunology; primary immunodeficiency; hyper IgE syndrome; hereditary angioedema; hematopoietic stem cell transplantation; space immunology; HIV infection
Concern for potential adverse effects of antiretroviral (ARV) chemotherapy used to prevent mother-to-child HIV transmission has led the US Public Health Service to recommend long-term follow-up of ARV-exposed children. Nucleoside reverse transcriptase inhibitor ARV agents can inhibit DNA polymerase γ, impairing mitochondrial DNA (mtDNA) synthesis and resulting in depletion or dysfunction.
We measured the mtDNA content of stored peripheral blood mononuclear cells (PBMCs) of 411 healthy children who were born to HIV-uninfected women and 213 uninfected infants who were born to HIV-infected women with or without in utero and neonatal ARV exposure. Cryopreserved PBMC mtDNA was quantified by using the Primagen Retina Mitox assay.
Geometric mean PBMC mtDNA levels were lower at birth in infants who were born to HIV-infected women. Among HIV-exposed children, mtDNA levels were lowest in those who were not exposed to ARVs, higher in those with exposure to zidovudine alone, and higher still in those with combination nucleoside reverse transcriptase inhibitor exposure. A similar pattern was observed in the corresponding women. Levels of mtDNA increased during the first 5 years of life in all HIV-exposed children but achieved normal levels only in those with ARV exposure.
Levels of mtDNA are lower than normal in HIV-exposed children. Contrary to expectation, PBMC mtDNA levels are significantly higher in ARV-exposed, HIV-uninfected infants and their infected mothers compared with ARV-unexposed infants and women. By 5 years, levels of PBMC mtDNA rise to normal concentrations in ARV-exposed children but remain depressed in ARV-unexposed children.
HIV; mitochondria; antiretroviral agents
More than 20 North American academic centers account for the majority of hematopoietic stem cell transplantation (HCT) procedures for primary immunodeficiency diseases (PIDs), with smaller numbers performed at additional sites. Given the importance of a timely diagnosis of these rare diseases and the diversity of practice sites, there is a need for guidance as to best practices in management of patients with PIDs before, during, and in follow-up for definitive treatment. In this conference report of immune deficiency experts and HCT physicians who care for patients with PIDs, we present expert guidance for (1) PID diagnoses that are indications for HCT, including severe combined immunodeficiency disease (SCID), combined immunodeficiency disease, and other non-SCID diseases; (2) the critical importance of a high degree of suspicion of the primary care physician and timeliness of diagnosis for PIDs; (3) the need for rapid referral to an immune deficiency expert, center with experience in HCT, or both for patients with PIDs; (4) medical management of a child with suspicion of SCID/combined immunodeficiency disease while confirming the diagnosis, including infectious disease management and workup; (5) the posttransplantation follow-up visit schedule; (6) antimicrobial prophylaxis after transplantation, including gamma globulin administration; and (7) important indications for return to the transplantation center after discharge. Finally, we discuss the role of high-quality databases in treatment of PIDs and HCTas an element of the infrastructure that will be needed for productive multicenter clinical trials in these rare diseases.
Allogeneic hematopoietic stem cell transplantation; gene therapy; primary immunodeficiency; clinical trial
Previously, we presented evidence that at physiologic concentrations the green tea catechin, epigallocatechin gallate (EGCG), inhibited attachment of gp120 to the CD4 molecule on T cells, but the downstream effects of EGCG upon HIV-1 infectivity were not determined.
To evaluate the inhibition of HIV-1 infectivity by EGCG and begin preclinical development of EGCG as a possible therapy.
Peripheral blood mononuclear cells, CD4+ T cells, and macrophages were isolated from blood of HIV-1 uninfected donors. HIV-1 infectivity was assessed by an HIV-1 p24 enzyme linked immunoassay. Cell survival was assessed by cell viability by trypan blue exclusion assay; cell growth by thymidine incorporation; and apoptosis by flow cytometric analysis of Annexin-V binding.
EGCG inhibited HIV-1 infectivity on human CD4+ T cells and macrophages in a dose-dependent manner. At a physiologic concentration of 6μM, EGCG significantly inhibited HIV-1 p24 antigen production across a broad spectrum of both HIV-1 clinical isolates and laboratory-adapted subtypes [B (p<0.001), C, D, and G (p<0.01)]. The specificity of the EGCG-induced inhibition was substantiated by the failure of EGCG derivatives lacking galloyl and/or pyrogallol side groups to alter HIV-1 p24 levels. EGCG-induced inhibition of HV-1 infectivity was not due to cytotoxicity, cell growth inhibition, nor apoptosis.
We conclude that by preventing the attachment of HIV-1-gp120 to the CD4 molecule, EGCG inhibits HIV-1 infectivity. As this inhibition can be achieved at physiologic concentrations, the natural anti-HIV agent, EGCG, is a candidate as an alternative therapy in HIV-1 therapy.
EGCG; HIV; p24; subtypes; lymphocytes; alternative therapy; green tea; apoptosis; cytotoxicity; proliferation
The goal of HAART is to promote reconstitution of CD4+ T cells and other immune responses. We evaluated the extent and the kinetics of immune reconstitution in HIV-infected children over 144 weeks of successful HAART.
Thirty-seven children receiving their first HAART regimen had plasma HIV RNA; T cells and subpopulations; T-cell rearrangement excision circles (TREC) DNA; candida, HIVCD4 and HIVCD8 enzyme-linked immunospot measured at regular intervals.
Plasma HIV RNA became undetectable in 81% of patients at 24 weeks and remained undetectable in 77% at 144 weeks. In contrast, CD4+% continuously increased. Distribution of T-cell subpopulations changed rapidly during the first 48 weeks of HAART and more slowly thereafter. At 144 weeks, total, naive and activated CD4+% and naive CD8+% of HIV-infected children were not significantly different from those of healthy age-matched controls, whereas total and activated CD8+% remained elevated. CD4+ and CD8+ TREC content increased only during the first 48 weeks of HAART. They positively correlated with each other and with total CD4+%, naive CD4+% and naive CD8+%. Candida and HIVCD4 enzyme-linked immunospot increased over time reaching peak values at 48 weeks and 144 weeks, respectively. HIVCD8 enzyme-linked immunospot decreased in magnitude over 144 weeks of HAART but retained its breadth. Baseline CD4+% positively correlated with CD4+% and with functional immune reconstitution at week 144, whereas baseline TREC correlated with TREC at week 144.
HIV-infected children acquired normal distribution of CD4+ T cells and other subpopulations and recovered CD4-mediated HIV immunity after 144 weeks of HAART.
candida; cell-mediated immunity; children; highly active antiretroviral therapy; HIV; T-cell rearrangement excision circle; T-cell subpopulations
We report an 8-year-old boy with AIDS, extremely elevated serum immunoglobulin G (IgG) concentration and IgG kappa [IgG(κ)] and IgG lambda [IgG(λ)] paraproteinemia. This paraproteinemia partially responded to highly active antiretroviral therapy. This case emphasizes the importance of controlling B-cell activation.
Carcinoma of the cervix is causally related to infection with the human papillomavirus (HPV), and T cells play a pivotal role in the immune response of the host to rid itself of HPV infection. Therefore, we assessed the T-cell function of women with HPV-related cervical neoplasia against a superantigen, Staphylococcus enterotoxin B (SEB). Each woman provided a cervical brush specimen for HPV DNA testing and Papanicolaou (Pap) smears for the staging of cervical lesions. They also provided a blood specimen for determination of the ability of CD4+ T and CD8+ T cells to synthesize Th1 (interleukin-2 [IL-2], gamma interferon [IFN-γ], and tumor necrosis factor alpha [TNF-α]) and Th2 (IL-10) cytokines in response to activation with SEB. Compared with control subjects with self-attested negative Pap smears, women with high-grade squamous intraepithelial lesions (HSIL) had significantly lower percentages of activated CD4+ T cells that produced IL-2 (P = 0.045), IFN-γ (P = 0.040), and TNF-α (P = 0.015) and a significantly lower percentage of activated CD8+ T cells that produced IL-2 (P < 0.01). These data indicate that women with HPV-related cervical HSIL show a decrease in Th1 cytokine production by activated CD4+ T cells and suggested that compromised T-helper functions may negatively impact the function of cytotoxic CD8+ T cells.
Objective: The mechanism whereby the placental cells of a human immunodeficiency virus (HIV)-1-infected mother protect the fetus from HIV-1 infection is unclear. Interferons (IFNs) inhibit the replication of viruses by
acting at various stages of the life cycle and may play a role in protecting against vertical transmission of HIV-1. In
addition the β-chemokines RANTES (regulated on activation T cell expressed and secreted), macrophage inflammatory
protein-1-α (MIP-1α), and MIP-1β can block HIV-1 entry into cells by preventing the binding of the
macrophage-trophic HIV-1 strains to the coreceptorCCR5. In this study the production of IFNs and β-chemokines
by placental trophoblasts of HIV-1-infected women who were HIV-1 non-transmitters was examined.
Methods: Placental trophoblastic cells were isolated from 29 HIV-1-infected and 10 control subjects. Supernatants
of trophoblast cultures were tested for the production of IFNs and β-chemokines by enzyme
linked immunosorbent assay (ELISA). Additionally, HIV-1-gag and IFN-β transcripts were determined by a
semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) assay.
Results: All placental trophoblasts of HIV-1-infected women contained HIV-1-gag transcripts. There were no
statistical differences in the median constitutive levels of IFN-α and IFN-γ produced by trophoblasts of HIV-1-
infected and control subjects. In contrast, trophoblasts of HIV-1-infected women constitutively produced significantly
higher levels of IFN-β protein than trophoblasts of control subjects. Furthermore, the median levels of
β-chemokines produced by trophoblasts of HIV-infected and control women were similar.
Conclusions: Since there was no correlation between the placental HIV load and the production of interferons or
β-chemokines, the role of trophoblast-derived IFNs and β-chemokines in protecting the fetus from infection with
HIV-1 is not clear.
An external evaluation program for measuring the performance of laboratories testing for cytokines and immune activation markers in biological fluids was developed. Cytokines, chemokines, soluble cytokine receptors, and other soluble markers of immune activation (CSM) were measured in plasma from a healthy human immunodeficiency virus (HIV)-seronegative reference population and from HIV-seropositive individuals as well as in supernatant fluids from in vitro-stimulated human immune cells. The 14 components measured were tumor necrosis factor (TNF) alpha, gamma interferon, interleukin-1 (IL-1), IL-2, IL-4, IL-6, IL-10, Rantes, MIP-Ia, MIP-Iβ, soluble TNF receptor II, soluble IL-2 receptor alpha, β2-microglobulin, and neopterin. Twelve laboratories associated with the Adult and Pediatric AIDS Clinical Trial Groups participated in the study. The performance features that were evaluated included intralaboratory variability, interlaboratory variability, comparison of reagent sources, and ability to detect CSM in the plasma of normal subjects as well as the changes occurring in disease. The principal findings were as follows: (i) on initial testing, i.e., before participating in the program, laboratories frequently differed markedly in their analytic results; (ii) the quality of testing of a CSM in individual participating laboratories could be assessed; (iii) most commercial kits allowed distinction between normal and abnormal plasma CSM levels and between supernatants of stimulated and unstimulated cells; (iv) different sources of reagents and reference standards frequently provided different absolute values; (v) inexperienced laboratories can benefit from participating in the program; (vi) laboratory performance improved during active participation in the program; and (vii) comparability between analyses conducted at different sites can be ensured by an external proficiency testing program.
Nineteen children with human immunodeficiency virus (HIV) infection
were treated with recombinant human gamma interferon (rIFN-γ) (50
μg/m2 subcutaneously three times each week during weeks 1
through 12 and 100 μg/m2 subcutaneously three times each
week during weeks 13 through 24) in a phase I/II clinical trial. All
children continued to receive previously prescribed therapy with oral
zidovudine or didanosine. Children were assessed clinically and with
laboratory studies during 24 weeks of study treatment and for 12 weeks
after completion of rIFN-γ therapy. In general, rIFN-γ therapy was
well tolerated. There were two clinical or laboratory adverse events
thought to be possibly or probably study drug associated. One child
developed acute pancreatitis; another child developed granulocytopenia.
Median CD4+-lymphocyte counts and plasma HIV RNA
concentrations did not change significantly during therapy. In vitro
neutrophil bactericidal activity against Staphylococcus
aureus and superoxide production were not significantly affected
by rIFN-γ therapy. We conclude that rIFN-γ therapy in HIV-infected
children receiving single-agent antiretroviral therapy is safe and does
not produce consistent changes in CD4+-lymphocyte count,
plasma HIV RNA concentration, or in vitro neutrophil function.
Lautropia mirabilis, a pleomorphic, motile, gram-negative coccus, has been isolated from the oral cavities of 32 of 60 (53.3%) children infected with human immunodeficiency virus (HIV) and 3 of 25 (12.0%) HIV-uninfected controls; the association of L. mirabilis isolation with HIV infection is significant (P < 0.001). All children in the study, both HIV-infected children and controls, were born to HIV-infected mothers. The presence of this bacterium was not associated with clinical disease in these children. The HIV-infected children with L. mirabilis did not differ from the HIV-infected children without L. mirabilis in immunological status, clinical status, or systemic medications. The role of HIV infection itself or concomitant factors in the establishment of L. mirabilis in the oral cavity remains to be elucidated.
Peripheral blood lymphocytes were isolated from 9 patients with chronic lymphocytic leukemia (CLL) and 12 normal control donors. The cells were assayed for synthesis of DNA and poly-(adenosine diphosphate ribose) (poly[ADPR]) immediately after isolation and on successive days following their treatment with phytohemagglutinin (PHA). Two different techniques were used to measure DNA synthesis. In the standard technique, DNA synthesis was measured by incubating intact cells with [3H]deoxythymidine. In the new technique, the lymphocytes were first rendered permeable to nucleotides, then DNA synthesis was measured by incubating them with [3H]deoxythymidine triphosphate in the presence of deoxyATP, deoxyGTP, deoxyCTP, ATP, and Mg++. Both assays showed the anticipated rise in DNA synthesis after PHA stimulation of normal cells. PHA-stimulated lymphocytes from patients with CLL demonstrated low levels of DNA synthesis in both assay systems.
The initial levels of poly(ADPR) synthesis were greater in CLL lymphocytes than in normal cells. Studies with a T-cell-depleted population of normal cells showed the same activity for poly(ADPR) synthesis that was demonstrated by the original population of normal cells. PHA stimulation produced an increase in poly(ADPR) synthesis in both the normal and CLL cells. The increase in poly(ADPR) synthesis in normal cells was coincident with the increase in DNA synthesis. The increase in poly(ADPR) synthesis in the CLL cells was dissociated from the delayed and diminished increase in DNA synthesis. Thus, CLL cells have higher than normal initial levels of poly(ADPR) synthesis. Poly(ADPR) synthesis is dissociated from DNA synthesis in CLL cells whereas it varies directly with DNA synthesis in normal lymphocytes.
Interaction of microgram quantities of highly purified rabbit anti-TNP antibodies with TNP-substituted HeLa, HEp-2, and L cells caused an intense stimulation of radioactive nucleoside ([125I]UdR and [3H]TdR) uptake which was maximal 24–72 h after exposure of cells to antibody. The stimulation of nucleoside uptake and presumaly DNA synthesis was shown to be immuno logically mediated because unsubstituted cells were not stimulated by anti-TNP antibody, normal rabbit gamma globulin did not stimulate TNP-cells, and a hapten inhibitor, ε-DNP-lysine, prevented the stimulation of TNP-cells by anti-TNP antibody. These findings demonstrate that interaction of antibody with cell surface antigen can alter cell membrane transport, and possibly can enhance cell growth.