RU2147238C1 - Method of treatment of patient with bacterial infection - Google Patents

Abstract

FIELD: medicine. SUBSTANCE: invention proposes to treat blood cells for two stages. At the first stage leukocyte cells suspension is obtained and incubated with cytokine complex and erythrocyte mass is washed out with physiological solution twice. At the second stage leukocytes are mixed with optimal dose of antibiotic and washed out erythrocytes and recovered to patient after incubation for 20 min. Method can be used for treatment of patients with severe bacterial infection. EFFECT: increased concentration of antibiotic in inflammation focus, enhanced immunological response of body. 1 tbl, 3 ex

Description

 The invention relates to medicine, namely to methods of extracorporeal hemocorrection with the use of antibiotics and cytokines in the treatment of patients with severe and generalized forms of bacterial infection in surgical, obstetric and gynecological and pediatric practice.

 A known method (PCT N 89/04665, class A 61 K 37/02, 1989) for the treatment of infectious diseases in children caused by the main immunological insufficiency of cellular immune function in the body, in which the child is prescribed an immunotherapeutically effective effective amount of cytokine - interleukin 2 (IL-2).

 There is also a method of treatment using IL-2 for advanced bacterial infections (German patent N 3910011, class A 61 K 37/02, 1990). According to this method, the administration of the IL-2 cytokine intravenously, subcutaneously, intraperitoneally, intramuscularly leads to an increase in the nonspecific cellular response, which helps to reduce the number of bacteria in the peritoneal cavity and lower endotoxins in the blood plasma.

 There is also a known method (PCT N 94/04174, class A 61 K 37/02, 1992), in which natural xenogenic (sheep) cytokines are used to enhance the immune response to the pathogen.

 However, with parenteral use of cytokines, they are rapidly eliminated from the bloodstream. This necessitates the use of cytokines in high doses, which can cause pronounced adverse toxic reactions.

 One of the ways to reduce the toxic effect of cytokines is the extracorporeal method of their use (RU N 2098125 C1, class 6 A 61 K 38/00, 1994), which consists in taking blood from a patient, isolating mononuclear cells, culturing them with a cytokine - IL -2 and return to the patient.

 Known methods for treating bacterial infections (PCT N 93/07892, class A 61 K 37/02, 1991; VS NN 5523288, class A 61 K 38/17, 1994) based on the use of a composition of biologically active peptides enhancing cell permeability, and antibiotics.

 There is also a method of treating a surgical (bacterial) infection (ed. St. NN 1805390, class A 61 K 37/02, G 01 N 33/49, 1990), in which, in order to reduce complications, an antibiotic is introduced in the average therapeutic dose into the cell suspension obtained by plasmapheresis, and after a 15-20 minute incubation, the resulting suspension is reinfused to the patient. The authors in the additional studies cited showed that the use of this approach allows for targeted transport of the antibiotic to the site of inflammation, increase the concentration of the antibiotic in the inflammatory site by 4 to 5 times compared with parenteral administration of the drug, and reduce the course dose of the antibiotic [1].

 Isolated antibacterial therapy is characterized by a bactericidal / bacteriostatic effect on pathogenic microorganisms, but is not able to correct immune dysfunctions, and in some cases exacerbates the state of immunosuppression. In turn, the use of only cytokines in the treatment of bacterial infection, although it helps to enhance immunological reactivity, however, far from in all cases it allows to eliminate the pathogen.

 The objective of the invention is to expand the scope and increase the effectiveness of the method of treatment of bacterial infection.

 The problem is solved in that in the method of treating a surgical (bacterial) infection, which consists in obtaining a cell suspension by plasmapheresis, incubating it for 15-20 minutes with an average therapeutic dose of an antibiotic and reinfusion to a patient, the processing of blood cells is carried out in two stages, the first of which whole blood is obtained from the leukemia suspension and it is incubated with a complex of cytokines, and the erythrocyte mass is washed twice with physiological saline, in the second, the leukocytes are mixed with the optimal dose of antibiotic and erythrocyte washing and after the 20th intubation incubation is returned to the patient.

10 thousand units of heparin are introduced into a sterile vial, 250-300 ml of the patient’s venous blood are taken, 20% gelatinol solution is added and incubated for 45 min at 37 ^o C. The suspension is collected in a separate vial, centrifuged at 100 rpm and the supernatant is removed. Suspension cells are resuspended in phosphate buffered saline and incubated with a solution containing a complex of native cytokines. The erythrocytes remaining in the vial (after collection of the leukoprecipitate) are washed twice with physiological saline, mixed with the cells of the leukoprecipitate and the optimal dose of the antibiotic, incubated for 20 min at 37 ^o C and reinfused to the patient.

Clinical examples
Example 1. A man, 62 years old. He was admitted to the surgical infection department of the Municipal Clinical Hospital N 12, Novosibirsk on 10.30.97 with a diagnosis of an extensive purulent wound in the sacrum, osteomyelitis of the right femur, wound sepsis (St. aureus), multiple organ failure syndrome II degree (hepatitis, toxic anemia, myocarditis). Concomitant diagnosis: septic endocarditis.

On August 28, 1997, an operation was performed for osteosarcoma of the sacrum. In the postoperative period - suppuration of the postoperative wound, the formation of pressure sores in the region of the right greater trochanter, the development of wound sepsis and multiple organ failure. The patient was given antibacterial therapy (4 courses of parenteral administration of antibiotics of 3–4 generations), detoxification, infusion therapy, and plasmapheresis. Despite intensive therapy, the patient's condition remained serious: weakness, lethargy, pallor of the skin, sweating, poor appetite, weight loss of 18 kg, hepatosplenomogaly, hyperthermia up to 38.5 ^o C. In the blood - toxic granularity of neutrophils, toxic anemia, high ESR, urine distase, the mass of medium molecules - 0.76 srvc Secondary immunodeficiency with predominant damage to the T-cell and monocytic link: the absolute and relative contents of CD3 ⁺ T-lymphocytes, regular subpopulations of CD4 ⁺ and CD8 ⁺ T-cells, immunoregulatory CD4 / CD / index, the number of HLA-DR ⁺ activated monocytes, proliferative are reduced cell activity. Conducted therapy is ineffective. Assessment of the general condition of admission on the SAPS (Simplified Acute Physiology Score) scale is 19 points, the probability of death is 43%.

 Treatment with the proposed method was started on November 3, 1997. Four consecutive procedures were performed for incubating the patient’s fractionated blood with a mixture of natural cytokines and antibiotics in the optimal dose.

After completion of therapy according to the proposed method, the patient returned to normal body temperature, appetite, sleep, the patient became active, the wounds began to quickly clean and fill up with granulations. According to laboratory tests: anemia decreased, toxic granularity disappeared, blood amylases, urine diastasis, and the level of medium molecular peptides normalized. Since 11/05/97, croke crops are sterile. According to the immunogram: the absolute number of CD3 ⁺ T cells, the relative content of CD4 ⁺ T lymphocytes and HLA-DR ⁺ activated monocytes increased, the immunoregulatory index and proliferative activity of cells normalized.

 Subsequently, the patient adequately responded to all ongoing therapy. November 28, 1997 was discharged in satisfactory condition.

 Example 2. A woman, 32 years old, was treated in the intensive care unit of the Municipal Maternity Hospital No. 4 of Novosibirsk with a diagnosis of 3 days after a small caesarean section. Condition after preeclampsia. Multiple organ failure syndrome. Postoperative intestinal paresis. Endometritis of a severe degree. Endotoxemia. Secondary immunodeficiency.

Laboratory: anemia, leukocytosis, high ESR, leukocyte intoxication index (LII) - 10 units, the weight of medium molecules - 0.89 units Secondary immunodeficiency with a predominant lesion of the T-cell and monocytic link: the absolute and relative content of T-lymphocytes and their sub-fractions, the number of HLA-DR ⁺ activated monocytes are sharply reduced, the functional (proliferative) activity of the cells is sharply inhibited. Conducted therapy is ineffective. Assessment of the general condition on the SAPS scale is 22 points, the probability of death is 50%.

 The treatment with the proposed method was started on October 6, 1997. Five consecutive procedures of incubation of fractionated blood with a mixture of natural cytokines and an antibiotic in the optimal dose were performed.

As a result of therapy according to the proposed method, the following was recorded in a patient: breakage of a fever after the 3rd procedure; a decrease in the severity of endogenous intoxication (LII decreased to 2 units, the level of average molecular peptides decreased to 0.32 units); exit from a critical state; resolution of intestinal paresis; improvement of subjective well-being. According to immunological examination: an increase in absolute lymphocytosis; absolute and relative content of CD3 ⁺ T- and CD72 ⁺ B-lymphocytes; restoration of the normal level of proliferative activity of cells, the number and functional activity of monocytes. On 13.10.97, the patient was transferred from the intensive care unit to the postpartum ward and was discharged in satisfactory condition on 24.10.97.

Example 3. Girl, 1 year old. From 03.11.97, she was treated in the children's intensive care unit of the Regional Clinical Hospital in Novosibirsk with a diagnosis of right-sided polysegmental destructive pneumonia, bronchopleural fistula, and respiratory failure of the II-III degree. 2 courses of antibacterial therapy (cephalosporins, amikacin), sanitation bronchoscopy, infusion, detoxification therapy were carried out. Despite the treatment, the patient's condition remains serious, intoxication is increasing (LII 6.7 r. Units), the body temperature rises to 38.5 - 39 ^o C, chills, weakness, toxic anemia and toxic-hypoxic encephalopathy are noted. According to the immunogram: absolute and relative lymphopenia (the content of T-cells and their suyopopulations, B-lymphocytes is reduced), inversion of the immunoregulatory index, inhibition of the functional activity of lymphocytes and monocytes.

 Treatment with the proposed method was started on November 18, 1997. Four consecutive procedures of incubation of fractionated blood with a mixture of natural cytokines and antibiotics in the optimal dose (based on 1 kg of the child's weight) were performed.

At the end of therapy according to the proposed method, the patient showed positive radiological dynamics, normalized body temperature, obstruction of the bronchopleural fistula occurred, manifestations of respiratory failure significantly decreased, and the effects of brain hypoxia were completely stopped. According to the hemogram: normalization of the leukocyte formula, a decrease in ESR, restoration of the number of red blood cells, the concentration of hemoglobin. According to the immunogram: the absolute number of lymphocytes and CD3 ⁺ T cells increased, the ratio of CD4 / CD8 T lymphocytes normalized, and the proliferative activity of cells increased. 24.11.97, the patient was transferred from the intensive care unit and subsequently adequately responded to all therapy. 12/9/97, was discharged in satisfactory condition.

 The proposed method was used to treat 69 patients: 22 people with severe forms of surgical infection, 21 septic patients, 16 puerperas with purulent-septic complications in the subsequent period, and 10 resuscitation children with severe and destructive diseases of the bronchopulmonary apparatus.

The following are comparative data on the level of predicted and actual mortality in patients with severe forms of surgical infection (n = 22) during treatment according to the proposed method:
Indicator - Treatment by the proposed method
Leukocyte intoxication index - 5.9 ± 0.8 r.ed.

General condition for SAPS - 11 ± 0.7 points
The predicted mortality rate is 21 ± 2.0%.
Actual mortality - 0%
Of the entire group of treated patients (n = 67), 2 out of 19 septic patients died. Multicenter randomized trials of the European Society for Intensive Medicine (ESICM) have shown that mortality in sepsis over the past 10 years remains at 38–39.4% [2]. In the treatment of septic patients with the proposed method, the mortality rate was 9.5% (2/21).

 Thus, the use of the proposed method can significantly improve the treatment of patients with severe and generalized forms of bacterial infection in surgical, obstetric, gynecological and pediatric practice.

Literature
1. Lokhvitsky S.V., Gulyaev L.A., Zubtsov N.V., Klimakhin Yu.D., Shvetsov D.A. Clinical pharmacokinetics of antibiotics when administered in the cell mass during plasmapheresis. // Health of Kazakhstan. - 1992. - N 8. - S. 22 - 24.

 2. Bukumirovic V., Bumbasirevic V., Pantic J., et al. An analysis of mortality in intensive care. // 8th European Congress of Intensive Care Medicine. - V. 2 Free Papers (Ed. C. Roussos), Athens (Greece). - 1995 .-- P. 437-440.

Claims (1)

 A method of treating a bacterial infection, which consists in obtaining a cell suspension by plasmapheresis, incubating it for 15-20 minutes with an average therapeutic dose of an antibiotic and reinfusing the suspension obtained to a patient, characterized in that the processing of blood cells is carried out in two stages, the first of which is obtained from whole blood leukemia suspension and incubate it with a complex of cytokines, and the erythrocyte mass is washed 2 times with physiological saline, on the second the leukocytes are mixed with the optimal dose of antibiotic and washed red blood cells, and after 20 min tnoj incubation returned to the patient.