PPD-Induced Monocyte Chemotactic Factor Production by Human Milk Cells

The functional capability of antigen-stimulated breast milk cells to produce an immunologic mediator was examined. Colostrum and comparison peripheral blood samples were obtained from ten women, two to four days postpartum, and supernatants from PPD-stimulated mononuclear cell cultures were assayed for the lymphokine, monocyte chemotactic factor. Five of the ten women studied had a history of a positive tuberculin skin test and one had received BCG immunization. Peripheral blood lymphocyte cultures and colostrum cell cultures from four of these six women produced monocyte chemotactic factor. These results demonstrated the functional capability of antigen-stimulated colostra! cells to produce immunologic mediators.     

Human milk cell migration and production of monocyte chemotactic factor: lack of activity

The lymphocytes of human breast milk have been previously shown to be immunologically competent while the monocytes have been described as actively motile in some reports but not in another. Cells were separated from milk samples collected by lactating women 2 to 9 days postpartum. Milk cell migration and production of the lymphokine, monocyte chemotactic factor, by milk cells were assayed using membrane filters. Milk cells were poorly motile although prior culturing improved their migratory ability. Cell-free milk was chemotactic for blood monocytes. Chemotactic factor production was detected in only two of 16 phytohemagglutinin-stimulated milk cell cultures.     

Chemotactic defects in severe combined immunodeficiency.

Cellular and humoral components of leukotaxis were studied serially in four male infants with severe combined immunodeficiency disease. Two of the four, both lacking B and T cells initially, had a significant defect in neutrophil and monocyte chemotaxis. The other two, who had a high number of immunoglobulin-bearing cells (B cells), did not have these cellular abnormalities. It contrast, defective generation of chemotactic factor following endotoxin activation was observed in all patients. The defects were corrected coincident with or soon after successful engraftment of either bone marrow or fetal tissues. The reported deficiencies may be another manifestation of the heterogeneity in SCID.     

Comparative studies of functional characteristics of mononuclear cell subsets and granulocytes

Two human peripheral blood monocyte subsets and lymphocytes were isolated by counterflow centrifugal elutriation (CCE). The cell volumes of 303 mu3 and 380 mu3 were measured for the smaller and larger monocyte populations, respectively. Superoxide release by large monocytes exposed to opsonized zymosan was five times more active than that of the small monocytes. The production of colony stimulating activity was two-fold greater and the myeloperoxidase activity was 1.4-fold greater by the larger monocytes. Enriched fractions of cytotoxic cells responsible for natural killer (NK) activity against neuroblastoma cells were also obtained by counterflow centrifugal elutriation. Natural killer cells were obtained in larger lymphocyte fractions and had a mean cell volume of 180 mu3. Compared with the NK activity against the neuroblastoma cells, both the small and large monocytes displayed greater antibody-dependent cellular cytotoxicity (ADCC) activity against human erythrocytes. The larger peripheral blood monocytes aggregated in response to FMLP (N-formyl-methionyl-leucyl-phenyl alanine peptide) and PAF (platelet activating factor). Unlike the granulocyte, monocyte aggregation in response to FMLP was not accompanied by degranulation nor was it potentiated by cytochalasin B. In addition, monocyte aggregation could be blocked by benoxaprofen, unlike the granulocyte. Thus, CCE provides a means of isolating subsets of monocytes and lymphocytes and obtaining large numbers of peripheral blood monocytes for functional studies.     

Biochemical characteristics of "young" and "old" erythrocytes of the newborn infant.

Cord blood samples from ten term infants were fractionated into populations of young and old erythrocytes and compared with cells prepared in a similar fashion from eight normal adults. The old cell fraction from the newborn infants had very low phosphofructokinase activity and did not display the usual decline of activity of the enzymes phosphoglycerate kinase and enolase. In addition, the old cells from the newborn infants demonstrated impaired glucose consumption, which, upon analysis of the pattern of glycolytic intermediates, appeared to be a result of the phosphofructokinase deficiency. These findings suggest that cells produced earlier in gestation possess the developmental characteristics of fetal blood to a more significant degree and that these biochemical alterations may produce functional impairment.     

Characteristics of impaired chemotactic function in cord blood leukocytes.

Mobilities of cord blood granulocytes were studied using the agarose plate method and Boyden's chamber method. In the agarose plate, granulocytes of cord blood were shown to have moderately decreased responses in chemotaxis and chemokinesis induced by Escherichia coli-derived chemotactic factor and/or zymosan-activated serum, whereas they were shown to have a normal capacity of random mobility. Although their distance and index of chemotaxis or chemokinesis in the agarose plate were significantly less than those of adult granulocytes, response rate in both types of mobility were evidently higher compared with those in patients with chemotactic defect. Furthermore, there is a difference between chemotactic responses of cord blood granulocytes to E. coli-derived chemotactic factor and to zymosan-activated serum in the agarose plate method. Using the latter, a more distinguishable difference between chemotactic responses of cord blood granulocytes and adult granulocytes was shown. The Boyden's chamber method tended to show a more significant difference between chemotactic responses of granulocytes of cord blood and adults than in the agarose plate method.     

Ontogeny of the immune system: fetal lamb as a model

We examined adult sheep lymphocytes for the following surface markers: Surface membrane immunoglobulin, antigen identified by anti-sheep thymocyte globulin and complement receptors. We quantitated sheep peripheral blood mononuclear leukocyte proliferative responses to mitogens and alloantigens, and demonstrated a neutrophil-directed chemotactic factor (N-LDCF) liberated by mitogen-stimulated sheep mononuclear cells. A comparison of adult and fetal sheep from 120-150 d of gestation demonstrated that 1) fetal sheep had adult proportions of surface membrane Ig+ and T+ lymphocytes but a significant decrease in FcIgG receptor+ lymphocytes, 2) proliferative responses of fetal sheep blood mononuclear cells to mitogens and alloantigens were comparable to adult responses, and 3) fetal mononuclear leukocytes failed to produce the lymphokine, N-LDCF, in response to mitogen stimulation.     

Transient remission after intercurrent measles infection in a patient with hyperimmunoglobulin E syndrome

A 5-yr-old patient with hyper IgE syndrome contracted measles. This was accompanied by a temporary disappearance of his skin lesions. The patient had a long history of recurrent infections, chronic eczematoid pruritic dermatitis, and elevation of serum IgE level since infancy. Immunologic studies revealed decreased suppressor T cells (OKT8 cells) with increased IKT4/OKT8 ratio, defect in suppressor T cell function, and decreased chemotactic index. In February 1985, when he developed an interrcurrent measles infection at age of 5, the eczematoid pruritic dermatitis disappeared completely and immunologic defects improved transiently, with normalization of OKT4/OKT8 ratio, decrease in in vitro IgE synthesis, in vivo serum IgE level, and interleukin-2 production, decreases in IgG Fc receptor-bearing cells and autologous mixed lymphocyte reaction, and normalization of chemotactic index. One month later, the eczematoid skin lesion relapsed and immunologic defects reappeared. These studies suggested that the pathogenesis of hyper IgE syndrome involved a hypofunction of suppressor T cell. The transient remission associated with measles infection is probably related to the effect of the virus on the helper T cells, resulting in a normalization of OKT4/OKT8 ratio and IgE production.     

Effect of intralipid on the phagocytic and microbicidal capacity of human monocytes in culture

We studied the effect of Intralipid (IL) in monocyte cultures based on the ability of the cultures to phagocytose and kill Candida albicans and produce the oxidative burst. The IL was taken up by monocytes in cultures, and these cells phagocytosed more Candida organisms than did the control cells [85 +/- 2.2% in the IL treated (1%) compared to 68 +/- 2.3% after 1 h in the control]. The percentage of killing of Candida albicans, which had been taken up by the IL-treated monocytes measured after 2 h in culture (48.3 +/- 6.0%), was no different when compared to control (47.0 +/- 5.8%). Following ingestion of IL, there was an increase in basal H2O2 production, however, the presence of the IL in the cells had no effect on the expected increase in H2O2 production following stimulation with either phorbol myristate acetate (PMA) or zymosan particles. Compared to untreated cells, a significant increase in the number of monocytes with positive nitroblue tetrazolium staining was observed in monocytes that had ingested IL (when they were stimulated with either PMA or Candida microorganisms). Similar results were obtained in monocyte-derived macrophages (i.e., monocytes in monolayer cultures for 10 days). These findings suggest that the essential monocyte functions of phagocytosis, microbicidal activity, and ability to elicit an oxidative burst are not directly altered by the conventional use of IL in clinical practice.     

人脐血移植重症联合免疫缺陷小鼠的实验研究

为了探讨人脐血移植到小鼠体内的生长特性，将人脐血细胞注入经亚致死剂量照射后的严重联合免疫缺陷（ＳＣＩＤ）小鼠。１０只移植人脐血的小鼠中５只存活达２８天，其造血和部分免疫功能得到恢复。利用人粒－单系祖细胞培养和ＣＤ＋４５细胞流式细胞仪检测及人ＨＬＡ－ＤＰＢ基因聚合酶链反应－寡聚核苷酸单链构像多态性分析等方法，检测出了植入ＳＣＩＤ小鼠骨髓中的人脐血细胞。提示，人脐血细胞可以植入ＳＣＩＤ小鼠并重建其造血和免疫功能。     

Elevated production of interleukin-2 by lymphocytes from children with acute leukemia

Abnormalities of the production of interleukin-2 (IL-2) may play an important role in the immunologic dysfunction observed in pediatric leukemia patients. For an evaluation of the ability of lymphocytes from leukemic children to produce this cytokine, the production of IL-2 by mitogen-stimulated peripheral blood mononuclear cells was determined in children with acute leukemia at the time of diagnosis, during clinical remission, and at the time of relapse. Of 16 patients, 11 (69%) with either acute lymphoblastic leukemia or acute nonlymphoblastic leukemia at the time of diagnosis had IL-2 production levels above the highest level observed in control subjects, and all but one had values above the control mean. Three of five treated patients had elevated IL-2 production at the time of bone marrow relapse. In addition, of 37 patients examined during clinical remission (both during chemotherapy and after the completion of maintenance chemotherapy), five had IL-2 production values above the control range and four of these five patients subsequently had relapses, compared with only one relapse in the remaining 32 patients with normal or below-normal levels of IL-2 production. These results demonstrate an increased ability to produce IL-2 by many patients with acute leukemia, both at the time of diagnosis and at relapse. Elevated IL-2 production may represent an immunologic response to leukemic cells and in some patients may provide a marker for persistent leukemia.     

Effects of insulin-like growth factor-I deficiency and replacement therapy on the hematopoietic system in patients with Laron syndrome (primary growth hormone insensitivity)

BACKGROUND: Primary insulin-like growth factor-I (IGF-I) deficiencies, such as in Laron syndrome (LS), are a unique model in man to study the consequences resulting from defects in growth hormone (GH) signal transmission. OBJECTIVE: To assess retrospectively the effect of IGF-I deficiency and its therapy on the various cells of the hematopoietic system as reflected by peripheral blood counts. PATIENTS AND METHODS: Two groups of patients were studied. The first group consisted of 11 untreated patients with LS, seven males and four females, who were followed from childhood into adult age. Average age at the time of data analysis was 45.4 +/- 9.6 years. The second group included ten children with LS, six males and four females, who received IGF-I replacement therapy for an average period of 6 years, ranging in age from 0.9-11 years. The mean age at initiation of therapy was 6.9 +/- 4.28 years. Only the seven children treated for 5 years or more were included in the analysis. Data on blood counts were collected from the patients' charts. Blood samples were drawn at baseline, weekly during the first month, once a month during the first year, and once every 3 months thereafter. Statistical analysis of the change over time was performed using repeated measures ANOVA. RESULTS: Children with LS had red cell indices in the lower normal range and an elevated monocyte count. A statistically significant rise in red blood cell (RBC) indices was seen in children during IGF-I therapy: RBC rose from 4.66 x 10(6)/ml to 4.93 x 10(6)/ml (p = 0.011); hemoglobin from 11.55 g/dl to 13.01 g/dl (p < 0.001); hematocrit from 34.94% to 38.52% (p = 0.007), and mean corpuscular volume from 72.27 fl to 79.93 fl (p < 0.001). The platelet count diminished significantly during IGF-I therapy from 316 x 10(3)/ml to 219 x 10(3)/ml (p = 0.02), and the monocyte count from 0.74 x 10(3)/ml to 0.49 x 10(3)/ml (p < 0.001). CONCLUSIONS: The present investigation, the first of its kind in this syndrome, confirms that IGF-I has a strong stimulatory effect on erythropoiesis. In addition, IGF-I therapy had a reducing effect on monocytes and platelets, an effect not previously described. The mechanism by which IGF-I mediates these effects needs further elucidation.     

Cytogenetic Abnormalities in Childhwood Acute Lymphoblastic Leukemias

From the early 1950s to the mid 1970s all childhood acute lymphoblastic leukemias were treated according to an unique protocol. It became evident that some parameters had prognostic significance: age, white blood cells count, lymphoma syndrome, initial meningeal involvement, mediastinal mass, treatment efficacy, and more recently immune markers and cytogenetics. Multivariate analysis demonstrated that karyotype is one independent prognostic factor for survival even when age, white cell count, tumor burden, and immunologic parameters are considered.     

Production of lymphotoxin and tumor necrosis factor by human neonatal mononuclear cells

Lymphotoxin (LT) and tumor necrosis factor (TNF) are cytokines with many common biologic effects including antiviral activity and induction of fever and the acute phase response; despite common effects, they are molecularly distinct. Because neonates are unduly susceptible to viral infection and frequently fail to mount a febrile response to infection, we hypothesized that neonatal cells would produce less LT and TNF than adult cells. We analyzed LT and TNF production by blood mononuclear cells and purified T cells using Northern blot analysis to detect specific messenger ribonucleic acid and specific assays to detect LT and TNF protein in culture supernatants. Compared to LT, TNF messenger ribonucleic acid and protein were produced more rapidly both by total mononuclear cells and by T cells in response to mitogen stimulation. Although there was intersubject variability, adult and neonatal mononuclear cells and T cells (n = 6) produced similar amounts of LT and TNF messenger ribonucleic acid and protein with similar kinetics. In experiments with phytohemagglutinin-stimulated mononuclear cells from ten additional subjects, supernatant LT was somewhat greater in neonatal cultures (neonatal = 62.8 +/- 60.5, adult = 13.2 +/- 10.7 units/ml, p less than 0.05), and TNF was somewhat greater in adult cultures (neonatal = 708 +/- 429, adult = 1987 +/- 392 pg/ml, p less than 0.01) at 24 h; results at 48 h and 72 h were similar. Thus, neonatal MC produced as much or more LT than did adult MC. Although the decreased production of TNF by neonatal MC was statistically significant, these cells did produce substantial amounts of this cytokine.(ABSTRACT TRUNCATED AT 250 WORDS)     

Prediction of persistent immunodeficiency in the DiGeorge anomaly

To assess the natural history of the immune defect in DiGeorge anomaly, we reviewed serial immunologic studies in 18 patients. The diagnosis was made with criteria based on the concept of the DiGeorge anomaly as a field defect. Initial or early follow-up laboratory examination suggested moderate to normal T cell function in 14 patients. None of these patients have lost T cell capability; they have never had infections characteristic of T cell deficiency. Four patients had clinical and laboratory evidence of profound immunodeficiency. A decreased number of CD4+ cells (less than 400/microliters) and a decrease in phytohemagglutinin responsiveness (stimulation index less than 10) may be useful in discriminating patients with immunodeficiency; absolute lymphocyte count and immunoglobulin values were not informative. At the time of surgery, the thymus was not found in 11 of 14 patients; however, only two of these patients had immunodeficiency. Patients with a persistently low number of CD4+ cells and decreased phytohemagglutinin response are candidates for immunologic reconstitution.     

Failure of allogeneic bone marrow transplantation to benefit HIV infection

A 16 year old boy underwent allogeneic bone marrow transplantation (BMT) from an human leukocyte antigen (HLA)-identical sibling for severe aplastic anaemia. He was symptomatic for 7 years before transplantation and had received multiple red blood cell and platelet transfusions. Conditioning for BMT consisted of cyclophosphamide, antilymphocyte globulin and total lymphoid irradiation. Engraftment was rapid, there was no evidence of rejection despite the history of multiple blood product transfusions and he did not develop acute or chronic graft versus host disease. He was well for the first 8 months after transplantation but then developed fevers, interstitial pneumonia, herpes simplex infections and cytomegalovirus enteritis. Serological studies revealed antibodies to human immunodeficiency virus (HIV) and he was considered to have acquired immune deficiency syndrome (AIDS). Retrospective analysis of the serum samples showed that he was seronegative for HIV until approximately 10 months before transplantation when his serum became HIV positive. Lymphocyte function studies done after transplantation suggested immunologic recovery at 3 months post-transplant with a brisk though subnormal response to phytohaemagglutinin stimulation. T cell subset analysis performed subsequently showed complete absence of CD4 positive cells indicating immune incompetence which was associated with clinical features of AIDS. Bone marrow transplantation had failed to produce sustained immunologic reconstitution and prevent the progression of HIV to which he ultimately succumbed.     

Improving the outcome of infants born at <30 weeks' gestation - a randomized controlled trial of preventative care at home

Background

Reference values for hematological and biochemical assays in pregnant women and in newborn infants are based primarily on Caucasian populations. Normative data are limited for populations in sub-Saharan Africa, especially comparing women with and without HIV infection, and comparing infants with and without HIV infection or HIV exposure.

Methods

We determined HIV status and selected hematological and biochemical measurements in women at 20–24 weeks and at 36 weeks gestation, and in infants at birth and 4–6 weeks of age. All were recruited within a randomized clinical trial of antibiotics to prevent chorioamnionitis-associated mother-to-child transmission of HIV (HPTN024). We report nearly complete laboratory data on 2,292 HIV-infected and 367 HIV-uninfected pregnant African women who were representative of the public clinics from which the women were recruited. Nearly all the HIV-infected mothers received nevirapine prophylaxis at the time of labor, as did their infants after birth (always within 72 hours of birth, but typically within just a few hours at the four study sites in Malawi (2 sites), Tanzania, and Zambia.

Results

HIV-infected pregnant women had lower red blood cell counts, hemoglobin, hematocrit, and white blood cell counts than HIV-uninfected women. Platelet and monocyte counts were higher among HIV-infected women at both time points. At the 4–6-week visit, HIV-infected infants had lower hemoglobin, hematocrit and white blood cell counts than uninfected infants. Platelet counts were lower in HIV-infected infants than HIV-uninfected infants, both at birth and at 4–6 weeks of age. At 4–6 weeks, HIV-infected infants had higher alanine aminotransferase measures than uninfected infants.

Conclusion

Normative data in pregnant African women and their newborn infants are needed to guide the large-scale HIV care and treatment programs being scaled up throughout the continent. These laboratory measures will help interpret clinical data and assist in patient monitoring in a sub-Saharan Africa context.

Trial Registration

nicalTrials.gov Identifier NCT00021671.     

Monocyte functional capacity in chronic neutropenia.

The bactericidal activity of monocytes from a child with chronic benign granulocytopenia who has had virtual absence of neutrophils yet minimal infections since birth was examined against Escherichia coli and Staphylococcus aureus and compared with that of monocyte and neutrophils from 20 control subjects. Studies on monocyte function in this patient with no neutrophils revealed normal monocyte kill of both organisms when compared with control monocytes. Monocyte and neutrophil killing of both organisms was similar in control subjects at bacteria to phagocyte ratios of 1:1. When ratios of 3:1 were employed, however, control neutrophils were more effective than control and patient monocytes in reducing the number of viable organisms. These findings support the neutrophil as the more effective blood phagocyte but stress the importance of monocyte functional capacity in patients compromised by granulocytopenia or neutrophil functional defects.     

A trial of alemtuzumab adjunctive therapy in allogeneic hematopoietic cell transplantation with minimal conditioning for severe combined immunodeficiency

For infants with SCID the ideal conditioning regimen before allogeneic HCT would omit cytotoxic chemotherapy to minimize short‐ and long‐term complications. We performed a prospective pilot trial with alemtuzumab monotherapy to overcome NK‐cell mediated immunologic barriers to engraftment. We enrolled four patients who received CD34‐selected haploidentical cells, two of whom failed to engraft donor T cells. The two patients who engrafted had delayed T‐cell reconstitution, despite rapid clearance of circulating alemtuzumab. Although well‐tolerated, alemtuzumab failed to overcome immunologic barriers to donor engraftment. Furthermore, alemtuzumab may slow T‐cell development in patients with SCID in the setting of a T‐cell depleted graft.     

Group B streptococcal cellulitis in infants: a disease modified by prior antibiotic therapy or hospitalization?

Among 78 cases of group B streptococcal infections in children hospitalized at our institution during 1981 to 1985, five cases of cellulitis in infants were identified. Age at onset was 4 to 11 weeks. Group B streptococci were isolated from each of two aspirate cultures, all four blood cultures obtained before administering antibiotics, and none of four spinal fluid cultures obtained. All five infants had previously been treated with IV antibiotics in the hospital. Two infants had been previously treated for group B streptococcal infections (one each meningitis and neonatal sepsis). In contrast, among the 29 other patients with late-onset (2 weeks of age or older) group B streptococcal infection, four had prior treatment with IV antibiotics (P less than .001). These data suggest that hospitalization and/or parenteral antibiotic therapy may be a risk factor for development of group B streptococcal cellulitis.