RU2377994C1 - Cancer immunorehabilitaiton mode - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention concerns medicine, namely oncology and can be used for cancer immunorehabilitation. According to the invention, the method implies the hypodermic implantation of porous titanic carrier incubators with the vaccine deposited with their pores. The porous titanic carrier incubators are used as said carriers, while the vaccine is presented with proper lymphokine-activated killers modified by the electromagnetic field.

EFFECT: invention allows lowering metastatic activity of a tumour owing to creation in an organism of functionally active immunocompetent cell depots ensuring activation of various immunity links within constitutive synthesis of cytokines.

2 cl, 1 ex

Description

The invention relates to medicine, namely to methods for immunorehabilitation of cancer.

The incidence of malignant neoplasms in recent decades in Russia is constantly increasing and, as most experts predict, the increase in the number of cancer patients in the 21st century will constantly increase. At the present stage of development of medical science, surgery and chemoradiotherapy remain the main methods of cancer treatment. It is known that the result of this therapeutic approach is the development of immunosuppression in the patient, which is often the cause of postoperative complications. In this regard, the need for the inclusion of biotherapy as a full-fledged component of the combined therapeutic approach in oncological practice is currently not in doubt among most specialists. The use of this method in the adjuvant mode after radical operations, chemo- and radiation treatments allows you to extend the time of a relapse-free period of life, improve the quality of life of patients. One of the main and most valuable areas of biotherapy in practical terms is the activation of the cellular link of antitumor immunity. For this purpose, methods of the so-called adoptive (from "adoptive" - introduced) immunotherapy are used. The essence of these methods is the activation of immunocompetent cells outside the body (in vitro) under the influence of cytokines. As you know, the main role in antitumor protection is played by natural killers (NK) and cytotoxic lymphocytes (CTLs). For most tumors, specific tumor antigens were not found, and many of them do not express MHC, and therefore CTLs cannot be recognized. NKs belong to the system of nonspecific immunity and, unlike CTLs, do not need MHC expression and the presence of antigens for the effective lysis of tumor cells. However, their number in the body is small, only 10-15% of all blood lymphocytes, which does not allow NK to cope with a significant tumor mass. Adaptive immunotherapy methods make it possible to obtain a significant amount of the so-called lymphokine-activated killers (LAC) from the total inactive fractions of the patient’s mononuclear lymphocytes in the blood. LAC is introduced into the patient's body, where they have a pronounced antitumor effect. LAC immunotherapy expands the spectrum of antitumor therapy possibilities. In addition, it has advantages over traditional methods: lack of toxicity, satisfactory tolerance of treatment, the possibility of use in conjunction with other traditional methods of treatment; in cases of drug resistance - stimulation of local antitumor cellular immunity, leading to tumor lysis, as well as improving the quality and longevity of patients. The LAC-therapy method is universal, since it can be used in almost any nosological forms of malignant neoplasms (melanoma, lung cancer, ovarian cancer, stomach cancer, etc.). The effectiveness of LAC in the treatment of cancer is a well-known fact. There is evidence of a positive experience with the use of LAC-therapy in the treatment of people (children and adults) with chemoresistant widespread malignant solid tumors. However, clinical studies have shown that adoptive immunotherapy is most effective in patients with malignant effusions (tumor pleurisy, ascites and pericarditis). According to the data, the clinical efficacy was 88%. It should be noted that the slowdown or cessation of fluid accumulation occurred only in those cases when a significant amount of activated lymphoid cells was noted in the pleural effusion. The study showed the effectiveness of the proposed method of adaptive immunotherapy for ascites in patients with disseminated ovarian cancer. During LAC-therapy, degradation and subsequent disappearance of the tumor cells from the malignant effusion of the cells was observed against the background of an increase in the number of activated lymphoid elements in 87% of patients, as well as a decrease in the tumor marker level (CA-125) in 50% and a partial regression of the tumor in 13% of patients. It was found that adoptive immunotherapy can have a normalizing effect on the immune status of cancer patients and contribute to the regression of tumor pleurisy and ascites, even with chemoresistant forms of ovarian cancer. In addition, at present, adoptive immunotherapy using LAC cells is mainly used to treat the so-called immunosensitive forms of malignant neoplasms: melanoma and kidney cancer. Unfortunately, along with the therapeutic effect, the authors sometimes note the appearance of negative side effects of this type of therapy. Patients experienced fever, chills, arthralgia, myalgia, nausea, vomiting, diarrhea, impaired vascular permeability, impaired respiratory function, and signs of liver damage. The most likely cause of such systemic reactions is the body's response to simultaneous, massive localized administration of a high concentration of cytokines and, above all, interleukin-2 (IL-2). It is logical to assume that in order to avoid such a negative effect of LAC-therapy, the preferred method is to create activated cells in the tissue depot that synthesize cytokines in the course of their life, which, getting into the blood, will be distributed throughout the body without creating a high toxic concentration in one local place. The structural basis for the creation of such a depot can serve as porous titanium carriers for cell structures (see patent RU 2300345, publ. 06/10/2007), which are the product of scaffold technology (from the English “scaffold” - scaffold from scaffolding), which are currently time is widely used in orthopedic practice.

As effectors in biotherapy, not only LAC, but also other immunocompetent cells are used. A variety of ways to increase their activity are offered. In particular, a very promising direction is the effect on cells of electromagnetic radiation. Despite the existence of different points of view on the primacy of certain physicochemical mechanisms of the influence of the electromagnetic field on biological objects, most researchers are unanimous that the field of this nature primarily affects the physicochemical processes and, through them, the rate of biochemical metabolic reactions by increasing the fluctuations of the charge components of ionic homeostasis that are able to respond to a given field frequency, the EMF changes the free energy of the system of charges of homeostasis (Paul OV AN "The impact of electromagnetic radiation on life". - M .: Helios ARV, 2002, p. 224). This assumption is consistent with the position put forward by L.D. Landau, who states that the free interaction energy of the system interacting with the field, depending on the correlation function of the biological macroscopic system, will change significantly (Landau L.D., Lifshits L.M. “Statistical Physics”. - M.: Gostekhizdat, 1958, p. 400). This in turn will lead to changes in the processes of cell division and growth, in particular, as a result of enzymatic reactions.

For various enzymes, EMFs with the same characteristics can both activate and suppress the corresponding reactions (see patent RU 2179578, publ. 02.20.2002). To date, a large amount of data has accumulated in the scientific literature, indicating a significant and many-sided effect on the prokaryotic and eukaryotic cells of wave factors of various origins. It was found that the resulting effect of the wave factor is due to both the physical characteristics of the exciting wave and the biological characteristics of the target cell, as well as the physicochemical characteristics of the interaction medium. So, it was proved that trace elements ions in minimal concentrations carry out both stimulation and inhibition of cell metabolism (Moralev L.N. et al. “Reactive cell changes under the influence of zinc biocomplex.” Advances in modern science. - Russian Academy of Natural Sciences, 7, 2007).

Closest to the proposed method is a method of treating cancer, which consists in the subcutaneous implantation of porous nickelide titanium carrier incubators with a vaccine deposited in their pores from EMF-treated biopsy tumor material (see patent RU 2285548, publ. 10.20.2006).

The disadvantages of this method are the use of porous nickelide titanium as carriers, since the nickel ions that make up the carriers, even at minimal concentrations, cause the development of cancer. The use of vaccines from "killed" tumor cells is dangerous in terms of passive antigenic stimulation of tumor growth.

The objective of the invention is to eliminate these disadvantages, the exclusion of the progression of the underlying disease and prolong life in cancer patients. The technical result consists in reducing the metastatic activity of the tumor, prolonging the patient's life and improving its quality.

The problem is solved, and the technical result is achieved by the fact that the method of immunorehabilitation of cancer is that they perform subcutaneous implantation of porous titanium carrier-incubators with a vaccine deposited in their pores, and their own modified lymphocytic cells are used as a vaccine. In order to prepare a vaccine from the patient’s own modified lymphocytic cells, peripheral blood puncture is performed.

The method is as follows.

Under local anesthesia with a 0.5% solution of novocaine in an amount of 30.0 ml, porous titanium structures (scaffold) containing modified own lymphocytic cells taken by peripheral vein puncture are implanted subcutaneously in the abdomen with a trocar.

Experimental studies were carried out on the basis of the Research Institute of ED and TO GU RONTs im. N.N. Blokhina RAMS. The aim of the work was to study the possibility of using titanium plates (scaffolds) as the structural basis of depot of effector immunocompetent cells as a promising method of immune biotherapy, as well as to assess the prospects of using autonomous electric pulse generators (AES GIT) to stimulate the effectiveness of immunocompetent cells.

In the study of immunomodulating activity in vitro, porous titanium plates (porosity 55-60%) having the shape of parallelepipeds (15 × 3 × 3 mm) were used as scaffolds. Dendritic cells and LAC were used as effectors.

The conducted experimental studies have established that the use of a titanium scaffold as a concentrating effector cell structure allows them to maintain viability and even slightly increase the cytolytic antitumor effect. This effect is probably due to the cytokines TNFα and IL-1β, synthesized by LAK and having a cytopathogenic effect on tumor cells. Since the high level of activity of such a cell colony is self-sustaining due to the synthesis of INFγ cells, it was assumed that the level of this cytokine in the medium should also be increased. However, we were not able to confirm this assumption. However, it should be noted that the applied cytokine analysis method allows only free mediator molecules to be identified. Based on this, it can be assumed that the cells synthesize INFγ in sufficient quantity to maintain their functional activity, but it quickly binds to the receptors of the producer cell itself or close to it and therefore is not detected in the environment surrounding the scaffold. Similarly, this argument can try to explain the detected low concentration of other cytokines. In this regard, the fact of an increase in the concentration of IL-5 in a medium containing a scaffold with LAC is extremely interesting. In a medium containing tumor cells in addition to the loaded scaffold, this cytokine cannot be detected. It is known that this cytokine enhances the expression of IL-2 receptors on the cell membrane, accelerating proliferation. Perhaps, in a concentrated arrangement, the molecules of this cytokine quickly bind to receptors, resulting in the long-term maintenance of a significant number of activated cells in the depot.

The obtained results allowed us to confirm the effect of direct cytotoxic effects of zinc ions under the influence of EMF, previously described by a number of researchers. We have shown that there is a differentiation of the effect of such an effect. So, tumor cells are 3 times more than LAC sensitive to its cytopathogenic effect, and the inhibition of respiratory activity of dendritic cells after exposure to iontophoresis was least pronounced - their IL in 3.2. times less than IL LAC.

Summarizing the foregoing, it can be argued that the introduction of scaffolds loaded into LAC into the body will contribute to the creation of a colony of functionally active immunocompetent cells in the body of the depot, which will ensure the activation of various parts of the immune system through the constant synthesis of cytokines.

The data obtained in the experiment made it possible to apply this method in the clinic.

Clinical example

Patient Sh., 43 years old, medical history 3294, was undergoing treatment at the Central Clinical Hospital of the Russian Academy of Sciences, Moscow, from September 10, 2007 to April 30, 2008 with a morphologically confirmed IHC diagnosis: Ring-shaped gastric cancer of the fourth stage, metastases Kruckenberg, peritoneal carcinomatosis, umbilical ring metastasis, inguinal lymph nodes, resistant ascites.

Upon admission, she complained of weight loss, anarexia, severe weakness. A clinical examination confirmed the diagnosis. The patient completed the first stage of immunorehabilitation. For the next three months, the patient was outpatiently monitored every 2 weeks and tested. Laparocentesis was performed according to indications within 10-12 liters of ascitic fluid. Three months later, the patient again entered the 2nd stage of immunorehabilitation. During the examination, there was an improvement in well-being, appetite, a decrease in ascitic fluid to 6 liters, a decrease and defragmentation of the inguinal lymph nodes. During the control EFGDS with targeted biopsy, the sclerotizing process of the proximal stomach tumor was detected without disturbing the motor-evacuation function of the stomach. On CT and ultrasound of the pelvic organs and ovaries, the process of formation of connective tissue of metastases of the ovaries and cysts in the pelvis was found. Monitoring of immunological parameters showed a growth trend in natural blood killers and interleukin-2. The next three months of outpatient monitoring showed steady stabilization of the main process and normalization of blood-immunological parameters. In April 2008, the patient entered the 3rd stage of rehabilitation, where after the examination all the indications for surgical treatment were found - cytoreductive removal of Kruckenberg metastases. On April 21, under endotracheal anesthesia, laparoscopy was performed, where it was found that the tumor-like formations of the ovaries softened, became mobile, cystic formations appeared on them and in places of metastasis in the parietal peritoneum. The process was recognized as operable, the conversion to lower-median laparotomy was performed and the ovaries were removed without technical difficulties, the abdominal cavity was drained, signs of invasive tumor growth were found, the abdominal cavity was sutured tightly.

Thus, based on experimental data, clinical observation data, the claimed method allows you to control the course of the tumor process and thereby extend and create comfortable living conditions for cancer patients.

Claims (2)

1. A method for immunorehabilitation of cancer, which consists in subcutaneous implantation of porous incubator carriers with a vaccine deposited in their pores, characterized in that porous titanium incubator carriers are used as such carriers, and their own lymphokine-activated killers modified by an electromagnetic field are used as a vaccine . 2. The method according to claim 1, characterized in that for the preparation of the vaccine using blood obtained by puncture of a peripheral vein.