Dynamics of the accumulation of the basic immunoglobulin classes in the intestinal contents in food toxinfections]

The work presents the data on the dynamics of accululation of the main classes of immunoglobulins (A, G and M) in coprofiltrates obtained from patients with alimentary toxicoinfections (bacteriologically confirmed salmonellosis and diseases of unknown etiology). The levels of immunoglobulins of all classes (mainly IgA and IgG) were shown to be elevated in the process of the disease. The dynamics of the increase in the level of IgA (both general and secretory) was supposed to indicate the formation of local immunity in the intestinal wall. The presence of serum IgA and the characteristic IgG dynamics seemed to be indicative of destructive processes occurring in the intestinal wall. Thus, the dynamics of accumulation of immunoglobulins in coprofiltrates obtained from patients with alimentary toxicoinfections reflects the main local pathological and immunological processes.     

Use of loading doses of bifidumbacterin forte for treatment of patients with acute enteric infections

The clinical observation of patients with acute enteric infections (AEI), treated with loading doses of Bifidumbacterin forte during the first 2 days of the disease, was carried out. The preparation was shown to produce a positive effect on the course of AEI: salmonellosis, alimentary toxicoinfections, acute, dysentery. The early decrease of the manifestations of intoxication, pain syndrome, diarrhea, as well as the acceleration of convalescence in comparison with standard treatment, were noted. The most essential dynamics was registered in salmonellosis patients. The analysis of clinical results allowed to recommend the use of loading doses of Bifidumbacterin forte, a probiotic with high colonization potential to normalize the microbiocenosis of the intestine in AEI.     

Cellular immunity factors in salmonellosis and food toxinfections

A complex immunological examination of 68 salmonellosis patients and 227 patients with alimentary toxinfections of unknown etiology has been made in the process of their treatment by rehydration therapy. At the acute period of the disease, irrespective of its nosologic form, cellular immunity has been suppressed. Cell-mediated immunity factors have been shown to depend on the severity of the disease.     

Effect of plasmids for multiple antibiotic resistance on the change in Salmonella typhimurium phagovar

The data on the influence of acquired plasmid resistance to antibiotics on S. typhimurium phagovar. Plasmid R was transferred from S. typhimurium strain, isolated from the focus of hospital salmonellosis and resistant to the lytic action of phages, to Escherichia coli K12 and then to antibiotic-sensitive S. typhimurium strains of different phagovars, isolated from patients with alimentary toxicoinfections. The influence of plasmid R on the phagovar of recipient strains, most pronounced in strains of phagovar I, was revealed. The transconjugates of this phagovar considerably changed sensitivity to phages and in some cases acquired resistance to the lytic action of typing phages, thus becoming identical in this feature to the donor of the plasmid.     

Improvement of the serological diagnosis of food poisoning]

A possibility was shown of improvement of serological diagnosis of salmonelloses with the aid of polyvalent (complex) and group salmonellosis stable formalinized erythrocytic diagnostic agents. The use of the mentioned preparations for titration of antibodies of different immunochemical nature in the sera of patients suffering from food toxicoinfections permitted to confirm the clinical diagnosis of salmonellosis in about 30% of cases.     

Synthesis and immunological effect of thymic humoral factor-gamma-2 analogues

Nine analogues of thymic humoral factor (THF)-gamma-2 were prepared by the solid-phase method, and their in vitro restoring effect on the impaired blastogenic response of phytohemagglutinin(PHA)-stimulated T-lymphocytes of uremic patients with infectious diseases were examined. The results were as follows: [Arg6]-THF-gamma-2 exhibited higher restoring activity than that of our synthetic THF-gamma-2. [Sar4]-, [Val1]-, [Arg3]-, [Gly5]-, and [Asn3]-THF-gamma-2 were also active but less potent than that of our synthetic THF-gamma-2. Three other peptides, [beta-Ala4]-, [Arg2]-, and [Gln2]-THF-gamma-2, did not show any restoring activity on the impaired blastogenic response of uremic patients with infectious disease.     

Synthesis and immunological effect of thymic humoral factor-gamma 2 analogues

Nine analogues of thymic humoral factor (THF)-gamma 2 were prepared by the solid-phase method and their in vitro restoring effect on the impaired blastogenic response of phytohemagglutinin(PHA)-stimulated T-lymphocytes of uremic patients with infectious diseases were examined. The results were as follows: [Arg6]-THF-gamma 2 exhibited higher restoring activity than that of our synthetic THF-gamma 2. [Sar4]-, [Val1]-, [Arg3]-, [Gly5]-, and [Asn3]-THF-gamma 2 were also active, but less potent than that of our synthetic THF-gamma 2. Three other peptides, [beta Ala4]-, [Arg2]-, and [Gln2]-THF-gamma 2, did not show any restoring activity on the impaired blastogenic response of uremic patients with infectious disease.     

Detection of tissue specific antigen-binding lymphocytes in patients with acute enteric infections

A total of 88 patients with salmonellosis, acute dysentery, alimentary toxicoinfection, acute gastroenterocolitis were examined. The study was aimed at early determination of the involvement of organs and tissues into the inflammatory process and detection of antigen-binding lymphocytes with the use of erythrocytic immunoreagents prepared from tissue antigens of mucous membranes of small and large intestine, duodenum, stomach, gall bladder, as well as liver and pancreas. The study demonstrated that as early as on day 1-3 of the disease the development of the inflammatory process in different organs was accompanied by the appearance of the corresponding tissue specific (organ specific) antigen-binding lymphocytes in all patients. As a rule, patients with different acute enteric diseases significantly differed by the frequency and spectrum of the involvement of such organs and tissues into pathological process.     

An immunohistochemical analysis of local immunity in Salmonella and dysenteric infections]

Immunohistochemical analysis was performed in 21 patients with acute and chronic dysentery, in 32 patients with different forms of salmonellosis in comparison with the cytoenzymatic status (CES) of immunocytes in the mucous membrane of the large intestine. It has been revealed that for acute dysentery the activation of the cellular and humoral links of local immunity is typical, but for salmonellosis--mainly of the humoral one. The chronic processes in dysentery and salmonellosis are connected with the increase in the subpopulation of T8-suppressors. Immunohistochemical data correlate completely with CES of immunocytes and that allows one use them with prognostic purposes.     

Melanin-based coloration and immunity in polymorphic population of pied flycatcher, <Emphasis Type="Italic">Ficedula hypoleuca</Emphasis>

A specific interest in the persistence of color polymorphism in some populations of birds and other vertebrates is often linked to ideas about the signaling honesty of bright coloration. The evolution of conspicuous ornamentation could be associated with physiological costs including limitations of the immune system. The study of this process is crucial for an understanding of the maintenance of polymorphic coloration. Here we summarized the results of a study of a pied flycatcher population from the Moscow region (Russia) in 2010–2013. We experimentally induced antibody production by injecting sheep red blood cells (SRBC) and inflammatory swelling by injecting phytohaemagglutinin (PHA) after which we estimated the immune response in breeding males. We used leucocytes-to-erythrocytes and heterophils-to-lymphocytes (H/L) ratios as indicators of infectious, inflammatory processes and stress. The results showed that the feeding rates of males treated with SRBC decreased and negatively related to the intensity of their immune responses. Non-molting males of different color types did not significantly differ in antibody production. Among molting breeders, the immune response to SRBC was significantly higher in pale males than in bright ones with rich melanin-based coloration. In contrast to non-molting males, molting pale males had an increased antibody titer after immunization. The lower humoral immune response was associated with the higher H/L stress index before immunization. The change in H/L after immunization positively correlated with the intensity of the humoral immune response. As opposed to humoral immunity, we did not find any significant predictors, including coloration, molt, or their two-way interaction, to explain the variation in cutaneous inflammatory response to PHA. The results suggest that the apparent advantage of a cryptic male phenotype over a conspicuous phenotype occurring in one of two types of immune response has an impact on the maintenance of color polymorphism in the pied flycatcher.     

Effects of interferon preparations on the course of Salmonella infection (experimental and clinical studies)]

The effect of type I interferons on the process of experimental salmonellosis in mice and rabbits, as well as their effect on salmonellosis in patients was studied. It was shown that homologous interferon increased the animal survival rate, activated bactericidal activity of the phagocytic cells and increased elimination of the infectious agents from the host. The complex of the etiotropic and pathogenetic treatment of 32 patients with salmonellosis included human leukocytic interferon, leukinferon. It was administered intramuscularly in a dose of 1 x 10(4)-2 x 10(4) IU: three injections at intervals of 48 to 72 hours (the treatment course). The course was repeated 10 days after the last injection of the first course. Addition of leukinferon to the routine scheme of the medicinal treatment of salmonellosis provided recovery of all the patients without complications or appearance of the bacteria carriers (the observation period of more than 3 years).     

Use of the indirect hemagglutination reaction for detecting antibodies to Pseudomonas pyocyanea in acute suppurative destructive pneumonia]

No less than 4-fold increase in the antibody titres to Pseudomonas aeruginosa during the infectious process served as laboratory confirmation of its participation in the infectious process. In 49 of 91 patients hemagglutining titre exceeded the diagnostic one (1:640). In 9 patients (chiefly in infants) hemagglutinin titre remained low, but there was a rise of antibody level to Pseudomonas aeruginosa during the disease. High hemagglutinin titres were noted in 12 patients on admission to the clinic, with reduction of the antibody titres at the late periods of the disease. Antibody titres remained unchanged during the disease in 15 patients. In 3 cases the indirect hemagglutination test was assesed as negative. In the rest of the patients hemagglutinin titres varied within the range of the diagnostic titre. Thus, the indirect hemagglutination test with erythrocytic diagnostic agent permitting to determine antibodies to Pseudomonas aeruginosa could be used at the clinic in combination with bacteriological and other investigations for establishing etiology of the destructive process.     

Clinical importance of infective agent persistence in Salmonella infection in children

The persistence properties of 70 S. enteriditis cultures and 12 S. typhimurium cultures isolated from the feces of children of the earliest age groups with the gastrointestinal form of Salmonellosis infection were studied. The study revealed that the low level of persistence properties occurred in most cases of mild forms of the disease. Salmonellae with a high level of anticomplement and antilysozyme activity were more often isolated in cases of moderate and severe forms of salmonellosis. The presence of correlation between the expression of the factors under study and the severity of the symptoms of intoxication in combination with the duration of the diarrhea syndrome in moderate and severe forms of Salmonella infection in children of the earliest age groups was established. The suggestion was made that the evaluation of the persistence properties of bacteria may be used for the prognosis of the course of the infectious process.     

A comparative evaluation of the sensitivity of different methods for the serological diagnosis of leptospirosis

In this work the comparative evaluation of the sensitivity and serological specificity of the microcapsular agglutination (MCA) test, the passive hemagglutination (PHA) test and the microagglutination (MA) test are presented. In the MCA test leptospiral antigens, adsorbed on synthetic carrier capsules produced by Japan Lyophilization Laboratory, were used and the PHA test was made with the use of polyvalent erythrocyte diagnosticum. The study of blood serum samples from 46 leptospirosis patients revealed that the values of antibody titers in the PHA and MCA tests were 5.5-8.1 times higher than the traditional MA test. In the MCA and PHA tests antileptospiral antibodies could be detected as early as on days 1-3 of the disease when the results of the MA test were negative or very low. The maximum values of antibody titers in the MCA and PHA tests were detected on days 11-15 of the disease and in the MA test, on days 21-25. The MCA and PHA tests are genus-specific and permit the detection of antileptospiral antibodies irrespective of the serogroup of the infective agent. In the study of the blood sera of 40 patients with diseases of nonleptospiral etiology the MCA and MA tests yielded false positive results in 7.5% and the PHA test, in 12.5% of cases in titers below the diagnostic level. These data are indicative of high sensitivity and specificity of the serological tests used in this study.     

Early results of the studies of the effect of cimetidine on selected parameters of humoral and cellular immunity in patients with peptic ulcer

The study aimed at evaluating an early effect of cimetidine on the blood IgG, IgA, IgM, number and functioning of T-cells in peripheral blood of patients with ulcerative disease. The study involved 27 patients (9 women and 8 men), aged between 27 and 70 years (mean 52.6 +/- 10.9 years). Nine of these patients suffered from the peptic ulcer and 8--from duodenal ulcer. Cell-mediated and humoral immunity were evaluated simultaneously in all patients prior to and after a 4-week treatment with cimetidine administered orally in the dose of 200 mg four times daily. Rosette test with theophylline and leukocyte migration test were used to assess cell-mediated immunity. It was found that cimetidine significantly increases blood serum IgG (p less than .01), the number of theophylline-resistant lymphocytes in TRFC-TR (P less than 0.01), and T-cell response to higher mitogen concentrations (PHA and Con-A) (p less than .005 and p less than .001, respectively).     

In vivo experimental modelling of the infectious process caused by stable L forms of the causative agent of typhoid

To simulate the infectious process and to study the persistence of L-forms, rabbits and guinea pigs were infected with S. typhi stable L-forms. The materials presented in this work indicate that both subconjunctival and intraperitoneal infection led to the development of the clinically indistinct, but morphologically pronounced pathological process with characteristic localization and typical changes in the gastrointestinal tract. The typical features of the process were the generalized immunomorphological reaction of the lymphoid apparatus with the appearance of light-colored reticulomacrophagal elements, the signs of the activation of humoral and cell-mediated immunity and the formation of small epitheloidocellular granulomas. The results of the investigation indicate that the stable cultures of S. typhi L-forms are highly pathogenic and capable of inducing the infectious process in experimental animals.     

Searching for biomarkers: humoral response profiling with luciferase immunoprecipitation systems

B-cell-mediated humoral responses are triggered in many human diseases, including autoimmune diseases, cancer, and neurologic and infectious diseases. However, the full exploitation of the information contained within a patient’s antibody repertoire for diagnosis, monitoring and even disease prediction has been limited due to the poor diagnostic performance of many immunoassay formats. We have developed luciferase immunoprecipitation systems (LIPS) that harnesses light-emitting proteins to generate high-definition antibody profiles that are optimal for both diagnostics and biomarker discovery. Here, we describe the results and implications from a range of LIPS-antibody profiling studies performed in our laboratory. These include highly sensitive diagnostics for domestic and global pathogens, insights into infection-related diseases, discovery of new biomarkers for human diseases, subcategorization of symptoms and identification of pathogenic autoantibodies against self-proteins. These investigations highlight the types of humoral response profiles associated with different diseases, provide new information related to disease pathogenesis and offer a framework for incorporating LIPS antibody profiling into global health initiatives and disease monitoring.     

Searching for biomarkers: humoral response profiling with luciferase immunoprecipitation systems

B-cell-mediated humoral responses are triggered in many human diseases, including autoimmune diseases, cancer, and neurologic and infectious diseases. However, the full exploitation of the information contained within a patient's antibody repertoire for diagnosis, monitoring and even disease prediction has been limited due to the poor diagnostic performance of many immunoassay formats. We have developed luciferase immunoprecipitation systems (LIPS) that harnesses light-emitting proteins to generate high-definition antibody profiles that are optimal for both diagnostics and biomarker discovery. Here, we describe the results and implications from a range of LIPS-antibody profiling studies performed in our laboratory. These include highly sensitive diagnostics for domestic and global pathogens, insights into infection-related diseases, discovery of new biomarkers for human diseases, subcategorization of symptoms and identification of pathogenic autoantibodies against self-proteins. These investigations highlight the types of humoral response profiles associated with different diseases, provide new information related to disease pathogenesis and offer a framework for incorporating LIPS antibody profiling into global health initiatives and disease monitoring.     

Renin-aldosterone response, urinary Na/K ratio and growth in pseudohypoaldosteronism patients with mutations in epithelial sodium channel (ENaC) subunit genes

Multi-system pseudohypoaldosteronism (PHA) is a rare syndrome of aldosterone unresponsiveness characterized by symptoms of severe salt-losing caused by mutations in one of the genes that encode alpha, beta or gamma subunit of epithelial sodium channels (ENaC). We examined long-term changes in the renin-aldosterone response in patients with different mutations. Four PHA patients were followed-up for 7-22 years. Patient A with a heterozygous Gly327Cys missense mutation in alphaENaC is a mild case and patients B, C and D are severe cases. Two additional patients with renal PHA served as controls. In patient A, serum aldosterone and plasma renin activity (PRA) decreased with age, PRA reaching near normal values at age 11. In contrast, patients B-D showed a positive correlation between age and aldosterone (r>0.86 for all). In patient B with Arg508 stop mutation, aldosterone reached 166nmol/L at age 19 (>300-fold higher than normal). Urinary Na/K ratios normalized gradually with age in all patients. Growth curves of the patients were reflective of the severity of PHA and compliance with salt therapy. Functional expression studies in oocytes showed that ENaC with alphaGly327Cys mutation, as observed in patient A, showed nearly 40% activity of the wild type ENaC. In contrast, stop mutation as in patient B reduces ENaC activity to less than 5% of the normal. Our results demonstrate distinct genotype-phenotype relationships in multi-system PHA patients. The degree of ENaC function impairment affects differently the renin-aldosterone system and urinary Na/K ratios. The differences observed are age-dependent and PHA form specific.