Nada Pejnović

Disseminated Bacillus Calmette-Guérin infection in a girl with hyperimmunoglobulin E syndrome.

Disseminated Bacillus Calmette‐Guérin infection occurs in few well‐defined immunodeficiencies, such as severe combined immunodeficiency, chronic granulomatous disease and paediatric acquired immunodeficiency syndrome. This severe complication of immunization against tuberculosis has been lethal in the majority of children who had primary immunodeficiency. Our patient, a 9‐y‐old girl with hyperimmunoglobulin E syndrome developed disseminated Bacillus Calmette‐Guerin infection in infancy. Patients with hyperimmunoglobulin E syndrome (HIES) are susceptible to serious staphylococcal and fungal infections. Disseminated Bacillus Calmette‐Guerin infection has not previously been reported in this rare immunodeficiency.

Interferon gamma alters the phenotype of rat thymic epithelial cells in culture and increases interleukin-6 production.

Rat thymic epithelial cells (TEC) in long-term culture were characterized by anticytokeratin monoclonal antibodies (mAbs) and electron microscopy. Phenotypic analysis performed by a large panel of mAbs showed that the highest percentage of these cells was of the subcapsular/medullary type. Recombinant rat interferon (IFN)-gamma up-regulated class-I and class-II MHC expression by TEC in culture as confirmed by immunohistochemistry and flow cytometry, but did not significantly alter other cell markers. TEC supernatants of IFN-gamma-treated cultures showed higher interleukin-6 (IL-6) activity, compared to the control, as determined by proliferation of the IL-6-sensitive B9-cell line. Increased IL-6 activity was probably not a consequence of increased TEC number in IFN-gamma-treated cultures because IFN did not significantly stimulate TEC proliferation in vitro. In contrast, IL-6 significantly stimulated TEC proliferation, indicating that this cytokine is not only a regulatory molecule for T-cell proliferation, but could also be an autocrine growth factor for thymic epithelium.

Long-Term Follow-Up and Prognosis of Chronic Granulomatous Disease in Yugoslavia: Is There a Role for Early Bone Marrow Transplantation?

We report the long-term follow-up of 12 pediatric-aged patients with chronic granulomatous disease (CGD). The mean age at the onset of infections was 5 months with a median delay in diagnosis of 2.5 years. Bacille Calmette–Guérin lymphadenitis was the most common presenting infection (6) followed by suppurative lymphadenitis (4), liver abscess (1), or Salmonella sepsis (1). Prophylaxis with cotrimoxazole was recommended to all patients. During the mean follow-up of 10 years (range, 4–23 years) pneumonitis was the most prevalent infection (91%) followed by lymphadenitis (83%), aphtous stomatitis (58%), and liver abscesses (25%). Seven (58%) patients developed chronic lung disease due to grossly delayed diagnosis (3) or poor compliance with antimicrobial prophylaxis (4). Five (41%) patients died during the second decade of life of aspergillosis (3) or chronic lung disease (2). Probability of survival into the third decade of life was estimated to be only 19%. We argue that HLA-identical bone marrow transplantation (BMT), if possible, should be attempted at early age because of significant morbidity and mortality in adolescence. BMT also should be considered in patients who suffer severe infections despite antimicrobial prophylaxis or patients with evidence of chronic lung disease. Possibility of elective BMT from unrelated donors remains to be carefully evaluated.

Post-traumatic activation of draining lymph node cells. II. Proliferative and phenotypic characteristics.

Proliferative and phenotypic characteristics of cells in regional lymph nodes that drain burn injury were examined in rats on day 3 postburn, i.e. at the time of maximal spontaneous proliferation and of interleukin-2 and accessory cytokine (IL-1 and IL-6) production. The importance of IL-1 in spontaneous proliferation of draining lymph node cells was demonstrated by stimulation of IL-2-driven proliferation by recombinant IL-1 in vitro and by susceptibility of unstimulated proliferation to anti-IL-1 antibodies, while requirements for IL-6 in draining lymph node cell proliferation were less pronounced. Cell surface phenotyping revealed a slightly increased percentage of CD25+ cells in the blast cell population of freshly isolated draining lymph node cells after injury, which increased further during cultivation. Enrichment in CD8+ cells on day 3 following burn injury was demonstrated, while no changes in total cell population and CD4+ cells was noted. This was however preceded by pronounced percentual decrease of total T cells and CD4+ cells and by increases of B cells and MHC class II+ cells on day 1 postburn. Inhibition of draining lymph node cell proliferation by anti-MHC class II antibodies suggested that this proliferation was class II MHC dependent. The contribution of cell proliferation and/or cell influx to day 3 postburn draining lymph node cell activity is discussed.

Increased activity of lymph node cells in experimental thermal injury: changes in accessory cells in injured area-draining lymph nodes

Accessory cell content and some of their functional characteristics were determined in regional lymph nodes which drain burn injury (DLN) in rats. Increase in percentages of non-specific esterase-positive cells and NBT+ macrophages and in numbers of dendritic cells were noted in cytospin preparations of draining lymph node cells (DLC) 24 and 72 h following thermal injury. An accumulation of B cells was also noted in the DLN paracortex region at these time points. Enrichment of ED1+ (rat macrophage marker) cells was noted in the adherent DLC population. Increased activity of interleukin-1 (IL-1) in conditioned medium from adherent DLC population and the increased stimulatory capacity of whole DLC or dendritic cell enriched-DLC fraction were noted in functional assays. Enrichment in accessory cells and an increase in their functional activity could contribute to the endogenous activity of regional lymph nodes which drain burned areas.

17 – An Excess of IL-6 Production in the Early Muscle Stage of Trichinella spiralis Infection in Mice is Associated with Strain Susceptibility to Infection

Publisher Summary This chapter presents a study that investigates the difference in IL-6 production between BALB/c (H-2d) and C57B1/6 (H-2b) strains of mouse infected with Trichinella spiralis (TS). The study was performed three weeks after infection, when the phenomenon of autoantibody synthesis amplification was observed and when the final stage of parasite development commenced, accompanied by muscle inflammation. Female BALB/c and C57B1/6 mice of 8–10 weeks of age and 19–22 g body weight were used in the experiments. Mice of each strain were randomly divided into two groups containing 10 animals per group. They were exposed to TS infection (group TS) and a control group (C) of nontreated mice. Mice were sacrificed on day 21 for the determination of muscle larva recovery, production of specific antibodies, and analysis of IL-6 cytokine production. Infectious L1 larvae were obtained by digestion of minced TS-infected rat carcasses in 1% pepsin-HCl for four hours at 37° C. The mice were infected by oesophageal intubation with 200 L1 larvae each. The results show that BALB/c mice harbored approximately 50% fewer L1 larvae in their muscles than C57B1/6 mice. This confirmed previous findings that BALB/c mice are considered to be resistant and C57B1/6 susceptible with respect to muscle larva burden. A significant increase of IL-6 production by the antigen-stimulated spleen cells was found in the T. spiralis susceptible C57B1/6 mice. The increase of IL-6 production in resistant BALB/c mice did not reach statistical significance when compared to the controls, uninfected mice.

IL-1, TNF and IL-6 Release by Wound- inflammatory Cells During the Healing Process in Two Strains of Rats

Publisher Summary This chapter presents a study in which the cellular responses during the course of wound healing in two different inbred rat strains, Albino Oxford (AO) and Dark August (DA), are investigated. The rate of wound closure of full-thickness wounds, the type of wound-infiltrating cells, and the time course of interleukin 1(IL-1), interleukin 6 (IL-6), and tumor necrosis factor (TNF) production by wound-derived cells were measured by using a sponge–matrix model. Two round full-thickness wounds were prepared dorsally in both strains with a sharp, round metal blade of 1 cm diameter, and wound diameters were measured during seven days of the healing process. The study demonstrates that DA and AO rats differ in their healing capabilities. The DA rats showed an accelerated wound-healing course that was accompanied by a different pattern of wound cellular infiltration and significantly higher levels of IL-6 and TNF in supernatants of wound-derived cells compared to AO strain. A higher proportion of infiltrating lymphocytes and higher IL-6 and TNF activities in wound-cell supernatants could be one of the mechanisms that underline the accelerated wound-healing course seen in DA rats.

Total Body Irradiation-induced Changes in Rat Serum IL-1, IL-6 and TNF Activities

Publisher Summary This chapter presents a study on the effects of various radiation doses on the rat serum interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor (TNF) activities. The study was performed by using male 8–10-week-old Albino Oxford (AO) strain rats, which were total body irradiated with 4 (LD10/30), 6.7 (LD50/30), 9 (LD95–100/30), and 12 Gy (LD100/30). IL-1 activity was measured by incorporating tritiated thymidine into IL-1-sensitive D10S cells derived from the murine T-helper cell line D10.G4.1. Serum IL-6 activity was measured by a B9 bioassay, and TNF activity was determined by an L929 fibroblast cell lytic assay. The specificity of the IL-1 and IL-6 assay was confirmed by using appropriate rabbit polyclonal anti-human IL-1 and IL-6 antibodies. The findings of this study showed that radiation doses ranging from a low lethal dose to an absolutely lethal dose caused the early increase (1–3 hours after irradiation) of the serum activity of IL-1 and IL-6, whereas the peak activity of TNF was found after 24 hours. At 6 hours and later, IL-1 activity in the sera of irradiated rats was significantly decreased. The second peak of IL-6 activity was found 3–5 days after irradiation. The study also demonstrated that IL-6 and dexamethasone produce the typical pattern of acute phase protein gene expression in the hepatoma cell line when given in combination.

27 – Altered Functions of Peripheral Blood Mononuclear Cells and Granulocytes in Patients with Active Psoriasis

Publisher Summary This chapter presents a study on the altered functions of peripheral blood (PB) mononuclear cells and granulocytes in patients with active psoriasis, such as the mitogen and phorbol ester-induced proliferative lymphocyte response, as well as neutrophil activation, adhesion, and tumor necrosis factor (TNF) production. The cells were isolated from venous blood of 16 patients (13 men and three women) with severe and active generalized forms of psoriasis, and six healthy volunteers were included as controls. Peripheral blood cells were isolated by centrifugation on a Lymhoprep density gradient, and the mononuclear cells were washed and resuspended in RPMI supplemented with 5% fetal calf serum. Statistical analyses were made by using the Mann–Whitney test. The study found that the level of spontaneous lymphocyte proliferation did not differ between psoriatic patients and healthy controls. Phytohaemaglutinin (PHA)-induced lymphocyte proliferation showed a significantly lower level in patients with psoriasis compared to healthy controls. Spontaneous neutrophil TNF production did not differ between psoriatic patients and healthy controls, except that four out of 16 patients with psoriasis had extremely high levels. The results of the study demonstrated impaired peripheral blood lymphocyte proliferative response and altered functions of PB neutrophils in patients with active psoriasis.

A Monoclonal Antibody R-MC 46 Induces Homotypic Adhesion and Activation of Rat Peripheral Blood Neutrophils

Publisher Summary This chapter presents a study in which homotypic adhesion is used to describe that R-MC 46 monoclonal antibody (mAb) triggers homotypic granulocyte aggregation and enhances phorbol ester (PMA)-induced NADPH oxidase activity and hydrogen peroxide production. Female Albino Oxford (AO) rats of 12–16 week old were used as a source of cells, and peripheral blood granulocytes were prepared by centrifugation on a gradient NycoPrep Animal 1007. The study found that the treatment of rat peripheral blood neutrophils by R-MC 46 mAb caused dramatic increase in aggregation, which was only partly dependent on β2 integrins and possibly involves other adhesion molecules. R-MC 46 two- to threefold enhanced the PMA-triggered hydrogen peroxide production and twofold enhanced nitroblue tetrazolium (NBT) reduction capacity. These findings suggest that the surface molecule R-MC 46 is the novel receptor for an activation pathway that leads in neutrophils to activation.