Treatment of chronic herpetic infection by vaccines based on IFN-induced dendritic cells - Causes, symptoms and treatment. MF.

One of the important problems of modern infectology is the chronic recurrent form of herpetic infection caused by the herpes simplex virus type 1 and type 2.The frequency of relapses directly depends on such a characteristic as the patient's quality of life. At the same time, the frequency of relapses 6 and more times a year with all the seemingly "harmless" infection has a significant impact on the overall health of the patient. According to the WHO Regional Office for Europe, herpes infection is defined as a group of diseases that determine the future of infectious diseases in the current century.

As it is known, the manifestations of chronic herpetic infection are quite specific and are manifested by single or grouped bubble elements with serous contents inside( vesicles) that appear against the background of "prodrome" - itching and unpleasant sensations in the place of future vesicles. The rash has its dynamic development: the formation in the dynamics of the on-site bubbles of crusts and subsequent hyperpigmentation. From the frequency of exacerbations of chronic infection depends on the form of the disease, and the quality of life of the patient. For one exacerbation an average of 1-2 weeks of recovery takes place, respectively, the capacity for work in this period is somewhat limited. Often the recurring form of infection is fixed at a frequency of exacerbations at least 4x6 times a year, even if the periods of remission last about 2.5 months. With a pronounced weakening of immunity, some patients complain of the frequency of exacerbations up to 15-20 times a year. Certainly, such patients should be selected individual and long-term therapy scheme using the latest methods.

Developments of herpes simplex

The developed therapy regimens for herpes infection include antiviral( acyclovir, valaciclovir, famciclovir and others) and immunomodulating( interferons, endogenous interferon inducers, immunotherapies of other groups) therapy that is effective enough at a certain stage of the disease course. Most therapy schemes are aimed at stunning the very exacerbation of the disease, but not aimed at long-term effect. This therapy is not always enough to achieve the desired result - reducing relapses to a minimum. A particular problem is the development of resistance( resistance) of viruses to antiviral drugs, especially used uncontrolled and without the appointment of a doctor. In this regard, scientific developments went further. The drugs and regimens of vaccine therapy for chronic herpetic infection have been developed. But these methods also lacked maximum activity. It was the turn to the newest scientific basis of treatment with the help of specific dentritic cells.

Dendritic cells of ( dendritic or DC) belong to the cells of the immune system and are a special kind of leukocytes having a branching structure( dendron-tree, Greek).Dendritic cells have a fairly large size - up to 15-20 microns and are located almost in all organs and tissues. Dendritic cells play an important role in the immune system - the function of representing the antigen. In other words, when a microorganism( viruses, bacteria, fungi, etc.) enters our body, the dendritic cells absorb it with the phagocytosis process, and then use its antigens to "represent" specific immunocompetent cells for antibody production( antigen-specific immune response).Dendritic cells are capable of activating with high strength such immunocompetent cells as CD4 and CD8 T-lymphocytes. However, many scientific studies have shown a significant reduction in the content of dendritic cells in patients with chronic viral diseases, which is considered as one of the reasons for the formation of a long persistence of viruses in the human body.

Dendrocytes

One of the modern methods for treating the chronic recurrent form of herpes infection is based precisely on the presentation by the dendritic cells of the antigen of the herpes virus in the presence of interferon-alpha and other components ["Method of immunotherapy for chronic recurrent herpesvirus infection" patent RU 2485962 on the basis of FGBU "ScientificResearch Institute of Clinical Immunology "Siberian Branch of the Russian Academy of Medical Sciences( FGBU" NIIKI "SB RAMS) Ostanin AA, Starostina NM, Chernykh ER].

The purpose of this method is to create a new generation of dendritic-cell vaccines that can solve one simple task - reducing the clinical manifestations of herpetic infection and reducing the frequency of relapses to a minimum with the help of an economically advantageous method of immunotherapy for chronic recurrent herpetic infection. The attainment of this goal is achieved by generating autologous interferon-alpha induced dendritic cells loaded with recombinant antigen of the herpes simplex virus in an in vitro culture from peripheral blood monocytes from the patient's peripheral blood monocytes, which the patient receives in 2 courses in combination with a recombinant interleukin-2 preparation as an adjuvantanswer).It is due to interleukin-2 is the activation of T cells, the processes of apoptosis( cell death) of lymphocytes, the synthesis of long-living cytotoxic T-lymphocytes are prevented.

The expected result of the therapy is the development of a single antigen-specific high-level immune response for a long period of time. Indications for treatment of

The indication for DC vaccination is a chronic herpetic infection, often a relapsing form with a relapse rate more than 6 times a year, and at least 2 weeks( that is, during remission) from the time of the last relapse.

How to treat dendritic cells

The technical implementation of the method is as follows: the patient is hospitalized in a hospital where the generation of dendrolettes from peripheral blood of the patient is performed. In the morning hours, 200 ml of blood is taken into a vial of heparin( 1000 U), to which is added a solution of gelatin( 40 ml) and incubation is carried out at 370 for 45 minutes. A leukocyte suspension is formed, which is centrifuged for 20 minutes at 1000 rpm. The precipitated cells are washed with phosphate buffered saline( PBS), layered on a density gradient of ficcolo-verographinum and again centrifuged for 20 minutes, but at 3000 rpm. The collected mononuclear cells( MNCs) undergo additional treatment( twice washed, resuspended) until OIC is isolated at a concentration of 3 * 106 / ml, which is incubated at 370 for 2 hours( this is the immature idlet).Subsequently, the cells undergo additional processing, the interferon-alpha load is 1000 U / ml( Roferon-A, Roche, Switzerland), repeated cultivation for 3 hours. Then, in the presence of the preparation polyoxidonium( NPO PetrovaxFarm, Russia) at a dose of 2 ng / ml and the final maturation of the dentritic cells takes place.

Generation of DK cells

In this way, IFN-DK( interferon-induced dendritic cells) is obtained, which are then loaded with the specific recombinant( artificially created in the laboratory) antigen of the herpes simplex virus( HSV 1 gD, NGO "Diagnostic Systems" Nizhny Novgorod) at a dosage of 5 μg / ml with incubation for 1 hour at 370.

The resulting preparation( specific IFN-DK) at a concentration of 6.0 * 106 / ml is cryopreserved for subsequent use. The entire manufacturing process lasts 4 days.

The method of DC-vaccination

The method of DC-vaccination with this drug consists of 2 courses of vaccine:

1) The "induction course" includes 4-6 subcutaneous injections in the upper third of the shoulder at a dose of 5,0 * 106 / ml at intervals of 2of the week. Vaccinations are carried out against the background of the simultaneous introduction of interleukin-2 - the drug "Roncoleukin ®"( OOO "Biotech", St. Petersburg) at a dose of 0.25 mg, which is administered subcutaneously as an adjuvant.

Injections 2) The "supportive course" consists of 3-6 subcutaneous injections of IFN-DK at a dose of 5 * 106 / ml at a frequency of once a month, simultaneously with adjuvant.

Control of the effectiveness of DC vaccination is carried out by determining the level of the proliferative response of MNCs to stimulation with the vaccinal antigen of the herpes virus( HSV 1gD) and mitogen( concanavalin A).The patient is taken 10 ml of venous blood, which is processed and within 72 hours the level of proliferation and other components of the study is assessed. The result is given in the form of an average score( cpm) of their three identical cultures. The control is carried out 4 times: before treatment, after 1 course, after 2 courses and 6 months after the end of therapy( last study 15 months after the start of treatment).

Treatment prognosis

Using this method of DK vaccination with IFN-DK vaccines revealed 75% efficacy in patients with a frequently recurring form of herpetic infection. Thus, the frequency of relapses in patients decreased by 3 times, and the total area of ​​lesions during exacerbation decreased, the duration of remission( inter-recessive periods) was extended to 3 and more months. There was also an improvement in the indices of a specific antigen-induced immune response both after the second course of vaccination and 6 months after the end of treatment.

Thus, we can talk about the emergence of a new effective method of DK vaccination, which can stimulate the development of an antigen-specific immune response, not only during vaccine therapy, but also 6 months after completion of treatment, which will undoubtedly affect the frequency of recurrences in this disease and the quality of lifepatients with chronic, often recurrent herpetic infection.

The doctor infektsionist Bykova N.I.