Tag Archives: recombinant human interleukin-7

Effect of recombinant human interleukin-7 on Pseudomonas aeruginosa wound infection

S. M. Grigorieva1, D. B. Starosyla1, S. L. Rybalko1,
V. V. Motronenko2, T. M. Lutsenko2,3, O. Yu. Galkin2

1Gromashevsky Institute of Epidemiology and Infectious Diseases,
National Academy of Medical Sciences of Ukraine, Kyiv;
2National Technical University of Ukraine “Igor Sikorsky Kyiv Polytechnic Institute”;
3UA Pro-Pharma LLC, Kyiv, Ukraine;
e-mail: alexfbt@gmail.com

Received: 01 July 2019; Accepted: 13 August 2019

A wide range of interleukin-7 (IL-7) biological effects suggests that application of appropriate preparations in clinical practice will stimulate immunity in patients with lymphocytic exhaustion or autoimmune diseases.  Studies are being conducted for IL-7 based preparations aimed at restoration of the immune system of patients with immunodeficiency of different origins. Pseudomonas aeruginosa is an important pathogen, which causes nosocomial infections in hospitalized patients. Infection factors, affecting the immune status of the host, play a key role. A promising and relevant scientific endeavor is the study of the effect of recombinant IL-7 (rIL-7) as an adjunct therapy in wound infections caused by P. aeruginosa. The aim of this study was to evaluate the effectiveness of rIL-7 use in P. aeruginosa wound infection in mice. The experiments were conducted using a standardized rIL-7 preparation, P. aeruginosa strain and 20 white non-inbred mice. The preparation of rIL-7 after all stages of purification was characterized by the content of ballast proteins and impurities via electrophoresis in a polyacrylamide gel in reducing conditions, and its biological activity was evaluated in the MTT test by means of proliferation of peripheral blood mononuclear cells. In the mice, the fur was removed, the neck nape was intentionally injured and P. aeruginosa bacteria were injected into the wound of each animal (0.1 ml of suspension with a bacterial cell concentration of 0.08×109 cells/ml). Starting from the 2nd day, bacterial examination of the wound material was carried out daily. Starting from the 3rd day, the mice (experimental group, n = 10) were intraperitoneally administered 5 μg (0.1 ml) of the rIL-7 preparation. In the control group of animals (n = 10), the rIL-7 preparation was not administered. In 80% of experimental animals (administered the rIL-7 preparation), the healing of wounds and elimination of the pathogen of purulent inflammatory infection P. aeruginosa occurred on the 7th day. On the 9th day from the beginning of wound infection, wound healing and elimination of P. aeruginosa occurred in all experimental mice. In 60% of mice from the control group (did not receive treatment with rIL-7), wound healing and the elimination of P. aeruginosa occurred on the 9th day. Wound healing and elimination of P. aeruginosa in all mice of the control group occurred on the 14th day. Thus, in mice treated with rIL-7, wound healing and elimination of the pathogen occurred 5 days earlier than in mice from the control group (without rIL-7 treatment). Subsequent studies may be aimed at developing protocols for the treatment of wound infections using an rIL-7 preparation in patients with a compromised immune system Therefore, rIL-7 is a promising preparation for the treatment of complex wound infections.

Development of the method for microbiological purity testing of recombinant human interleukin-7-based product

O. Yu. Galkin1, T. M. Lutsenko1,2, Yu. V. Gorshunov1, V. V. Motronenko1

1National Technical University of Ukraine
“Ihor Sikorskii Kyiv Polytechnic Institute”, Ukraine;
2UA “PRO-PHARMA” LLC, Kyiv, Ukraine;
e-mail: alexfbt@gmail.com

The goal of the work was justification of standardization parameters of the product based on recombinant human IL-7 by “microbiological purity” parameter and development of its measurement method with consideration of the requirements of guideline documents regarding quality of medicinal pro­ducts. Suitabi­lity assessment of microbiological purity testing procedures was performed. It has been established that the studied­ product in standard testing methods possessed antimicrobial activity against test microorganisms Basillus subtilis ATCC 6633 (on soybean casein digest agar and Sabouraud dextrose agar containing an antibiotic), Candida albicans АТСС 10231 (on soybean casein digest agar and Sabouraud dextrose agar containing an antibiotic), and possessed no antimicrobial activity against test microorganisms Staphylococcus aureus ATCC 6538 (on soybean casein digest agar) and Pseudomonas aeruginosa ATCC 9027 (on soybean casein digest agar). The efficacy of lecithin and polysorbate as substances eliminating antibacterial effect in combination with 50-fold dilution of the test sample has been suggested and proven for the purpose of antimicrobial activity neutralization. The developed total aerobic microbial count measurement procedure for the test product conforms to acceptance criteria provided by SPU (2.6.12), and can be used for its microbiological purity control.