INDEXES OF CYTOKINE PROFILE OF BLOOD IN PATIENTS WITH COMPLICATED ERYSIPELAS.

The cytokine blood profile in patients with complicated erysipelas was investigated. It was found that in patients with complications of erysipelas (gangrene, phlegmon, abscess, thrombophlebitis of the subcutaneous veins of the shin) levels of pro-inflammatory cytokines IL-1ß, TNF-α, IL-2, IL-6 in serum significantly increase and level of anti-inflammatory cytokine IL-4 increases slightly, as well as was found a significant increase in coefficients reflecting the ratio of pro-inflammatory and anti-inflammatory cytokines, which indicates the prevalence in the blood of examined patients with complications of erysipelas an anti-inflammatory properties. A more significant increase in pro-inflammatory cytokines serum levels is typical for patients with destructive forms of erysipelas - phlegmonous and gangrenous, a slight increase - for patients without purulent-necrotic component of complication (thrombophlebitis of the subcutaneous veins of the shin). In the future we plan to study pharmacological correction of shifts in cytokine blood profile with drugs with immunomodulating properties in patients with complicated erysipelas.

[INDICATORS OF PHAGOCYTIC ACTIVITY OF MONOCYTES AND CYTOKINE PROFILE IN PATIENTS WITH ERYSIPELAS OF VARYING SEVERITY].

Erysipelas (PB) ­ a serious disease that is accompanied by severe complications and high lethality. The immunological investigations were carry out in 51 patients who were treated at the clinic for erysipelas with an immunomodulator application. Marked changes in the indices of phagocytic activity of monocytes (PHAM) and the cytokine profile in patients with erysipelas were note. Application in complex treatment of immunomodulator give the positive effect on the indices of PHAM.

Acquisition of complement factor H is important for pathogenesis of Streptococcus pyogenes infections: evidence from bacterial in vitro survival and human genetic association.

Streptococcus pyogenes (or group A streptococcus [GAS]) is a major human pathogen causing infections, such as tonsillitis, erysipelas, and sepsis. Several GAS strains bind host complement regulator factor H (CFH) via its domain 7 and, thereby, evade complement attack and C3b-mediated opsonophagocytosis. Importance of CFH binding for survival of GAS has been poorly studied because removal of CFH from plasma or blood causes vigorous complement activation, and specific inhibitors of the interaction have not been available. In this study, we found that activation of human complement by different GAS strains (n = 38) correlated negatively with binding of CFH via its domains 5-7. The importance of acquisition of host CFH for survival of GAS in vitro was studied next by blocking the binding with recombinant CFH5-7 lacking the regulatory domains 1-4. Using this fragment in full human blood resulted in death or radically reduced multiplication of all of the studied CFH-binding GAS strains. To study the importance of CFH binding in vivo (i.e., for pathogenesis of streptococcal infections), we used our recent finding that GAS binding to CFH is diminished in vitro by polymorphism 402H, which is also associated with age-related macular degeneration. We showed that allele 402H is suggested to be associated with protection from erysipelas (n = 278) and streptococcal tonsillitis (n = 209) compared with controls (n = 455) (p < 0.05). Taken together, the bacterial in vitro survival data and human genetic association revealed that binding of CFH is important for pathogenesis of GAS infections and suggested that inhibition of CFH binding can be a novel therapeutic approach in GAS infections.

[Time course of changes in serum cytokine levels and the erysipelas clinical pattern].

The time course of changes in the values of proinflammatory tumor necrosis factor-alpha (TNF-alpha) and IL-8 and the anti-inflammatory cytokine IL-4 has been studied in 78 patients with erysipelas of different frequencies and pathomorphological forms of the disease. In erysipelas and its erythematous form, there is an increase in the levels of TNF-alpha and IL-8 in early phase of the disease, a decrease in these indices during convalescence, and a rise in the content of the cytokine IL-4, which are typical of an acute cyclic infectious process with a good outcome. In recurrent erysipelas and its most severe bullous-hemorrhagic form, there is imbalance in the production of cytokines that are suggestive of that there might be a disease recurrence. In erysipelas patients with a TNF-alpha value of 2.5 pg/ml or less at the onset of the disease, the relative risk for prolonged signs of systemic and local inflammatory reactions increases as compared with those who have a TNF-alpha level of more than 2.5 pg/ml.

[T-cell responsiveness to specific group A streptococcus antigens in patients with recurrent erysipelas].

Peripheral blood leukocytic migratory activity (LMA) was studied in 51 patients with recurrent erysipelas versus 63 patients with primary erysipelas. To reveal LMA, the authors employed in vitro a screening cell migration test as an indicator of the cooperation of T and B lymphocytes and macrophages, by stimulating with polysaccharide A, surface proteins, L-antigen, hyaluronidase, streptolysine-O, and a complete set of Grasse S. pyogenes. In patients with recurrent erysipelas, undulating LMA changes were found in the course of the disease in response to the stimulation with partial specific S. pyogenes antigens. There were differences in the time course of LMA changes, when stimulated with specific surface streptococcus antigens and with components of streptococcus with toxic activity. Significant LMA differences were found in relation to the pattern of a local process: active LMA changes from acceleration to suppression in the erythematous-hemorrhagic form and hyperergic reactions of LMA acceleration in the bullous-hemorrhagic one.

[Prognostic value of leukocytic migratory activity indices in patients with erysipelas].

Peripheral blood leukocytic migratory activity (LMA) was studied in patients with primary or recurrent erysipelas. A screening cell migration test (SCMT) was used in vitro and it established the prognostic value of MAL parameters at week 1 after the onset of erysipelas. It has been shown that a rapid transition of LMA from the phase of acceleration to that of inhibition characterizes the formation of an adequate immune response, corresponds to the good course of the disease, and has a low likelihood of recurrences. The probability of a recurrence is much higher than that when LMA tends to transit from suppression to acceleration and when LMA parameters are constant in the phase of suppression or acceleration. No transition of LMA to the phase of suppression in early convalescence suggests that the formation of an immune response to streptococcus is delayed.

[T-cell responsiveness to specific group A streptococcus antigens in patients with primary erysipelas].

Peripheral blood leukocytic migratory activity (LMA) was studied in 63 patients with primary erysipelas. To reveal LMA, a screening cell migration test (SCMT) was used as an indicator of the cooperation of T- and B-lymphocytes and macrophages in the stimulation with polysaccharide A, surface proteins, L-antigen, hyaluronidase, streptolysin O, a complete S. pyogenes antigen complex after Grasse. The prognostic value of MAL parameters was established at week 1 after the onset of erysipelas. A rapid transition of LMA from the phase of acceleration to that of inhibition was shown to characterize the formation of an adequate response, to correspond to the good course of the disease, and to be followed by the low likelihood of recurrences. The probability of a subsequent recurrence is much higher than that when LMA tends to transit from suppression to acceleration and when LMA parameters are constant in the phase of suppression or acceleration. No transition of LMA to the phase of suppression in early convalescence suggests that the formation of an immune response to streptococcus is delayed.

Analysis of Serum Cytokines and Single-Nucleotide Polymorphisms of SOD1, SOD2, and CAT in Erysipelas Patients.

Increased free radical production had been documented in group A (ß-hemolytic) streptococcus infection cases. Comparing 71 erysipelas patients to 55 age-matched healthy individuals, we sought for CAT, SOD1, and SOD2 single polymorphism mutation (SNPs) interactions with erysipelas' predisposition and serum cytokine levels in the acute and recovery phases of erysipelas infection. Whereas female patients had a higher predisposition to erysipelas, male patients were prone to having a facial localization of the infection. The presence of SOD1 G7958, SOD2 T2734, and CAT C262 alleles was linked to erysipelas' predisposition. T and C alleles of SOD2 T2734C individually were linked to patients with bullous and erythematous erysipelas, respectively. G and A alleles of SOD1 G7958A individually were associated with lower limbs and higher body part localizations of the infection, respectively. Serum levels of IL-1ß, CCL11, IL-2Rα, CXCL9, TRAIL, PDGF-BB, and CCL4 were associated with symptoms accompanying the infection, while IL-6, IL-9, IL-10, IL-13, IL-15, IL-17, G-CSF, and VEGF were associated with predisposition and recurrence of erysipelas. While variations of IL-1ß, IL-7, IL-8, IL-17, CCL5, and HGF were associated with the SOD2 T2734C SNP, variations of PDFG-BB and CCL2 were associated with the CAT C262T SNP.

Coley's toxins, tumor necrosis factor and cancer research: a historical perspective.

As far back as the 1700s, it was recorded that certain infectious disease processes could exert a beneficial therapeutic effect upon malignancy. Most prominent among the numerous deliberate efforts made to take advantage of these observations was that of a pioneering New York surgeon, William B. Coley, active career 1891-1936. Using a bacterial vaccine to treat primarily inoperable sarcoma. Coley accomplished a cure rate of better than 10%. This review examines the history of these efforts and presents a discussion of their corresponding relevance to present day immunotherapy.

Transient absence of C5a-specific neutrophil function in inflammatory disorders of the skin.

Chemotactic migration, production of superoxide anion (O2-), and the release of beta-glucuronidase from azurophilic granules were determined in polymorphonuclear leukocytes (PMN) from 135 patients with infectious (e.g., pyoderma, acne conglobata, erysipelas) as well as noninfectious (psoriasis) skin diseases. Purified C5a and the formylated tripeptide FMLP were used as stimuli. In addition, longitudinal profiles of PMN activities were performed at daily intervals in several patients. There was a complete absence of PMN responses (chemotaxis, O2--production, and enzyme release) specifically induced by C5a in 25 patients suffering from various inflammatory diseases of the skin. In these patients PMN responsiveness for the tripeptide FMLP was either normal or increased. The C5a-dependent defect of PMN was transient and correlated with disease activity. When normal PMN were incubated with sera from C5a-defective patients, no inherent stimulatory or inhibitory activities compared to control sera were seen. Pretreatment of normal PMN in vitro with various concentrations of C5a failed to completely deactivate PMN without affecting FMLP dependent functions. These observations demonstrate the presence of a functional defect in circulating PMN during acute cutaneous inflammation. The in vitro experiments suggest transient blocking of C5a-dependent PMN functions by a cell-bound factor which seems not to be C5a or C5adesarg.

Filariasis and erisipela in Santo Domingo.

This study examined acute-convalescent changes in diagnostic anti-streptococcal antibodies by the anti-streptolysin O (ASO) and anti-DNAase B (ADAB) tests among patients (n 28) with lymphedema and recurrent erisipela of the lower limb, comparing them with endemic normal control residents (n=25). The study was based in Villa Francisca, an urban focus of Bancroftian filariasis in eastern Santo Domingo, capital of the Dominican Republic. The acute signs and symptoms of erisipela were consistent with a diagnosis of bacterial cellulitis. The ASO test was especially successful at demonstrating a rise in mean titer during convalescence, whereas the ADAB produced about the same frequency of significant increases (0.2 log titer) as did the ASO. When subjects were scored as responders if mounting a minimal titer increase by either test, patients were found more frequently positive than were controls (chi2=5.3, P=0.02). About half (54%) of all patients mounted at least a minimal antibody increase. Filaria-specific IgG4 antibodies were absent from all sera of 20 residents of a nonendemic Dominican mountain town but appeared in about two-thirds of the sampled residents of the endemic barrio. Notably however, levels did not change between the acute phase and convalescence. These findings are consistent with the hypothesis that recurrent streptococcal invasion of the lymphatics may be a significant factor triggering or amplifying lymphedema and elephantiasis in patients with chronic filariasis.

Batch potency testing of inactivated erysipelas vaccines by ELISA--development, validation and implementation.

Inactivated erysipelas vaccines are widely used to protect pigs against erysipelas disease caused by the bacterium Erysipelothrix (E.) rhusiopathiae. Quality control tests for this vaccine are laid down in the European Pharmacopoeia (Ph.Eur.) Monograph No. 64. A laboratory animal model using a vaccination-challenge procedure is currently required as batch potency test. More than 10 years ago we initiated the first studies to develop an alternative ELISA potency model to replace this regulatory challenge test in mice. A short retrospective outline of the various steps from the development of the method until implementation into the regulatory requirements is described.

Potency testing of inactivated erysipelas vaccines by ELISA--influence of the adjuvant on antibody development.

The humoral immune response after vaccination may be greatly influenced by the type of adjuvant used. In this study we investigated the influence of three different adjuvants on the serological response in laboratory mice after immunisation with commercial erysipelas vaccines.

Regulating effects of porcine interleukin-6 gene and CpG motifs on immune responses to porcine trivalent vaccines in mice.

In order to develop novel immunoadjuvants to boost immune response of conventional vaccines, experiments were conducted to investigate the regulating effects of porcine interleukin-6 gene and CpG motifs as the molecular adjuvants on immune responses of mice that were co-inoculated with trivalent vaccines against Swine fever, the Pasteurellosis and Erysipelas suis. Synthetic oligodeoxynuleotides containing CpG motifs were ligated into pUC18, forming recombinant pUC18-CpG plasmid. Eukaryotic plasmid expressing porcine interleukin-6 (VPIL-6) were also constructed as molecular adjuvants in an attempt to enhance levels of immune responses of mice co-administered with the trivalent vaccines in this paper. The cellular and humoral immune responses of mice were systematically analysed, and the experimental results were observed that the number of white blood cells, monocytes, granuloytes and lymphocytes significantly increased, respectively, in the mice immunized with VPIL-6, compared with those of the control; the IgG content and titre of specific antibodies to the trivalent vaccine mounted remarkably in the sera from the VPIL-6 vaccinated mice; the proliferation of lymphocytes and induced IL-2 activities were significantly increased in the vaccinated groups. The above-mentioned immune responses of mice co-inoculated with pUC18-CpG plasmid were significantly stronger than those of co-inoculated with pUC18 plasmid, suggesting that the immunostimulatory effect of oligodeoxynuleotides CpG is closely connected with the number of CpG motifs. These results suggest that the porcine IL-6 gene and CpG motifs could be employed as effective immunoadjuvants to elevate immunity to conventional vaccines.

[Two cases of severe cutaneous vasculitis with thrombocytopenia associated with hepatic cirrhosis: autoimmune and infective-inflammatory component with endothelial lesion and restrictive pulmonary pattern in one case]. / Dois casos de vasculite cutânea grave com plaquetopenia associada à cirrose hepática: componente autoimune e infecto-inflamatório com lesão endotelial e padrão restritivo pulmonar em um caso.

Two clinical cases of female patients with hepatic cirrhosis and autoimmune multisystemic involvement with infectious intercurrent are reported. Case 1 presented infective endocarditis and erysipelas on the left thigh. In the course of the clinical picture a cutaneous vasculitis developed in the same place together with autoimmune thrombocytopenia, leukopenia and pulmonary restrictive picture with inflammatory pattern. There are also elevate immune complexes and complement consumption. Case 2 presented erysipelas on the left thigh cutaneous vasculitis and complement consumption. In Case 1 the infective endocarditis was treated with antibiotic therapy during 4 weeks followed by 1 mg/kg corticoid (Prednisone) with thrombocytopenia and leukopenia reversion. Case 2 presented an improvement with antibiotic only. The relation between chronic liver diseases and systemic autoimmune phenomena is commented, special attention being paid to the cutaneous, hematological and pulmonary affection.

[The functional characteristics of the peripheral blood granulocytes in erysipelatous inflammation]. / Osobennosti sostoianiia granulotsitov perifericheskoi krovi pri rozhistom vospalenii.

The receptors (FcR, C3R) and functional activity, determined by the nitroblue tetrazolium (NRT) test, of polymorphonuclear leukocytes (PML) of low and normal density were studied in erysipelas patients. The leukocytes were obtained by sedimentation on the 2-stage gradient of Ficoll-Verographin (1.077 and 1.119 g/cu cm). No statistically significant difference in the average group indices between "light" and "normal" PNL of erysipelas patients were detected. In comparison with donor PNL, higher expression of C3R, a high spontaneous NBT(+)-PNL level and poor response to stimulation with IgG in the NBT test were observed on granulocytes of the patients. The short-term treatment of the whole blood obtained from the patients with Streptococcus haemolyticus allergen led to a significant increase in the output of "light" PNL. As negative control, brucellin treatment was used, which produced no essential effect. The treatment of donor blood with the above-mentioned antigens did not significantly affect the density of PNL. These facts suggest that in erysipelas the presence of "light" PNL is linked not with the release of granulocytes from the marrow, but with the activation of leukocytes by the products of infective inflammation.

[Determination of antibodies against ribosomes and various cell wall components and detection of circulating antigens of group A Streptococcus in patients with erysipelas]. / Opredelenie antitel k ribosomam i nekotorym komponentam kletochnoi stenki i vyiavlenie tsirkuliruiushchikh antigenov streptokokka gruppy A u bol'nykh rozhei.

The level of antibodies to the ribosomes, polysaccharide A and peptidoglycan of group A streptococcus in the blood of patients with primary, secondary, and often relapsing erysipelas was studied by means of the enzyme immunoassay with the use of the sandwich techniques. For control, the sera of healthy donors were used. In the sera obtained from all groups of erysipelas patients a significant rise in the levels of antibodies to ribosomes and peptidoglycan in comparison with the controls was revealed. An increase in the level of antibodies to polysaccharide A was revealed only in patients with frequently relapsing and secondary erysipelas. Depending on the clinical form and the duration of the disease, polysaccharide A was detected in 32-51.9% of erysipelas patients and protein-ribosomal antigen was detected in 28.6-51.9% of such patients.