Extraction of Outer Membrane Proteins of Proteus Mirabilis Isolated From Urinary Tract Infections and their Immunological Effect In Vitro

Abstract

300 urine samples were collected from patients visiting Fallujah city hospitals (consulting clinics of Fallujah Teaching Hospital and Women and Children Hospital) for the period from 1/9/2020 to 30/12/2020, of different ages and for both sexes. The results showed that out of 300 urine samples were 244(81.33%) A sample with a positive result for bacterial culture, and it was found there were high significant differences between the positive and negative cultures. Females and males (64.70) (35.29), respectively.Isolates of Proteus mirabilis were diagnosed by observing the cultivar (colonies) and microscopic (bacterial cells) characteristics, biochemical tests and diagnostics with Vitek device. Antibiotic sensitivity were done at 12 types, Antibiotic resistance wre vary from Cefotaxime, Rifampicin, Ceftriaxon (100%), Cefixime, Ciprofloxacin (76.4%), Levofloxacin (64.7%), Nitrofurantoin (58.8%), Gentamicin (52.9%), Amikacin, Nalidixic acid (35.2%) (17.6%) Imipenem (11.7%).Extraction and partial purification of outer membrane proteins (OMPs) from the most antibioticresistant isolate using lysozyme, DNase, RNase, and N-Lauroyl-Sarcosinate enzymes. associated with the ion exchanger.To study the effect of the antigen of outer membrane proteins, some immunoassays were performed in vitro and the following results appeared: The use of concentrations (100, 50, 25) of OMPs antigen led to low significant of lymphocyte viability percentage (96.66, 97.66,98.0), respectively compared to With the negative control (99.66), the concentrations (100,50)% significantly reduced the percentage of survival of PMNs cells (97.66,98.33), respectively, compared with the control (99.66), while the concentration of 25 did not affect the survival of PMNs cells (99.66). The above concentrations showed a low significant in the diameter of the migration circuit of PMNs cells (8.830,10.59,12.21) compared with the negative control (15.21), while these concentrations led to a high significant in the percentage of formazone-forming PMNs cells (64.66,58.0,47.0) compared with the control (36.66). ) and a high significant in the sensitivity of lymphocytes (4.0,3.39,1.87), as the absorption spectrum values for concentrations (100,50,25) reached (0.92,0.74,0.41), respectively, compared with the negative control (0.22) and the positive control (1.53). The concentrations of the used proteins increased the phagocytic index (PI) of PMN cells ,The phagocytosis coefficient was high significantly with time until it reached its maximum at 90 minutes, and from the statistical analysis it was found that the phagocytosis coefficient increases with increasing concentration of proteins.