RU2485962C1 - Method for immunotherapy of chronic refractory herpes virus infection - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to clinical Immunology and Infectious diseases, and may be used for treating chronic refractory herpes virus infection. That is ensured by using autologous IFN-alpha induced dendritic cells loaded with recombinant herpes virus antigen and administered to the patient subcutaneously by two courses in a combination with a preparation of recombinant interleukin-2 as an adjuvant.

EFFECT: method enables reducing a number of recurrences and a severity of clinical manifestations of herpes virus infection ensured by induction of the effective antigen-specific cell-mediated response and maintaining it at an adequate level for a long period of time.

1 ex, 1 dwg

Description

The invention relates to medicine, namely to clinical immunology and infectious diseases, and can be used to treat chronic, often recurring herpes virus infection.

Herpes virus infection is one of the most important medical and social problems due to its wide spread and extremely large range of clinical manifestations and complications. Herpesvirus infection is caused by herpes simplex viruses type I and II, as well as the herpes virus of other varieties. According to a global review of herpesvirus studies, herpes infection / incidence is increasing from year to year. The variety of clinical manifestations, the characteristics of pathogens, the possibility of their spread by almost all known routes of transmission allowed the WHO Regional Office for Europe to attribute herpes virus infections to the group of diseases that determine the future of infectious diseases in the current century [1, 2]. It is important to note that according to modern epidemiological studies, not only an increase in the number of patients with herpetic infection, but also more severe forms of the course of the disease with frequent exacerbations is recorded [3].

In 85-90% of cases, the virus is under the control of the immune system. However, in 10-15% of people with weakened immunity, a clinical manifestation of the disease occurs, and the frequency of relapses can vary from 1-2 to 12-20 times a year. As a rule, herpes virus infection manifests itself in the form of rashes on the mucous membranes of the body (lips, nose, genitals), fever, chronic fatigue syndrome, and general depressive state. The clinical picture lasts from 5 days to 2 weeks, while the person is inoperative. In Russia, annually more than 2.5 million people undergo inpatient treatment in infectious, ophthalmological, neurological, and dermatological departments.

If herpes worsens more often than 1 time in 3 months, this indicates a chronic, often relapsing nature of the course of the disease, which is an indication for the appointment of adequate etiopathogenetic and immunomodulating therapy.

As a rule, for the treatment of chronic recurrent herpesvirus infection, antiviral drugs are used in combination with interferon inducer preparations (patent RU 2373951 C1) or with the preparation of recombinant human interferon 2-alpha (patent RU 2391981 C2), as well as with other immunomodulators, biostimulants, multivitamins and local drugs. The most widely used are acyclovir, valtrex, famciclovir, alpizarin, ribamidil, methisazone (antiviral drugs), as well as immunomodulators - ridostin, interlock, thymalin, lycopide, polyoxidonium, derinat, glutoxim, neovir, cycloferon, vitamins of group B. non-specific action and are not able to induce an antigen-specific immune response aimed at eliminating the virus. Clinical experience with the use of acyclovir or its analogues shows that antiherpetic agents can stop acute manifestations, but do not prevent the recurrence of chronic herpetic infection, and in some cases do not even reduce the frequency of relapses. In addition, there are no clear clinical and laboratory criteria for the appointment and sequence of use of certain combinations of various drugs. It is important to note that resistance is formed to most modern antiviral drugs, which also leads to relapse of the disease and a new cycle of therapy.

It is known that dendritic cells (DCs) capable of initiating an antigen-specific immune response play a key role in recognizing foreign (including viral) antigens and presenting them to effector T cells [4]. DCs have the unique ability to activate both CD4 and CD8 T cells. Moreover, the stimulating effect of DC is 10-100 times higher than the stimulating effect of macrophages and other antigen-presenting cells due to the higher level of expression of costimulatory molecules, slow intracellular degradation of antigen, and prolonged presentation of antigenic peptides on the cell membrane. In addition, DCs are the only antigen-presenting cells that can activate naive T cells [5].

A number of studies have shown that chronic infectious diseases of a viral nature are clearly associated with a decrease in the number and / or inhibition of the functional activity of DCs, which is considered as one of the possible mechanisms for the persistence of the virus in the patient’s body [6, 7].

Interest in the study of DC in viral diseases is associated not only with elucidation of their pathogenetic role in the chronicity / persistence of infection, but also with the potential use of DC for the development of new cellular technologies for the treatment of chronic and often recurring viral infections [8, 9], since DCs possess unique ability to induce an antigen-specific immune response.

The development of new approaches to the creation and use of therapeutic DC vaccines was made possible thanks to the development of methods for generating DC in an in vitro culture. According to standard protocols, DC generation is obtained by culturing an adherent fraction of peripheral blood mononuclear cells (CD14 + monocytes) in the presence of granulocyte macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) [10]. The immature dendritic cells (IL4-DC) generated in this way have a high ability to capture antigen, but weak stimulatory activity against T cells. Therefore, in the second stage, the final maturation of DC is initiated. Various factors are used for this: bacteria (living or dead), bacterial products (LPS), an analogue of viral RNA - polyinosine-cytidylic acid (poly-I: C), double-stranded DNA fragments, pro-inflammatory mediators and their combinations (IL-1 - beta , TNF-alpha, IL-6, prostaglandin-E2, CD40 ligand, etc.). At the stage of final maturation, DCs can be pulsed (loaded) with the “target” antigen against which an immune response must be induced. Such antigen can be, for example, lysates of tumor cells or bacteria, as well as immunogenic peptides of tumor or viral antigens (for example, obtained in the form of recombinant products). Mature DCs stably express on their surface molecules of the main histocompatibility complex (MHC class I and II) in combination with antigenic peptides, and also differ from immature DCs in increased expression of co-stimulatory (CD86) and adhesive (CD54, CD58) molecules, which contribute to a more effective interaction with T cells [11].

However, the use of GM-CSF for the generation of dendritic cells in combination with IL-4 is not the most optimal approach, since IL4-DC obtained in vitro have low migration activity and, under conditions of growth factor deficiency, can undergo reverse transformation into monocytes, and, accordingly, quickly lose their unique antigen-presenting properties.

Therefore, an alternative method for generating DC was proposed, in which IL-4 is replaced by interferon-alpha (IFN-alpha) [12]. This way of DC generation seems more physiological, since IFN-b is an early mediator of innate immunity and is produced in large quantities in response to stimulation by infectious antigens and pro-inflammatory cytokines. In addition, IFNb-induced DCs (IFN-DCs) have a number of features that fundamentally distinguish them from IL4-DCs and can significantly increase the efficiency of their clinical use. So, IFN-DK are generated faster in time, are characterized by high ability to capture antigen, maintain stability in the absence of cytokines and growth factors, have higher migration activity, more actively stimulate CD8 T lymphocytes, and induce a balanced cellular and humoral immune response, since they are capable of stimulate both Th1 and Th2 lymphocytes [13, 14]. Considering also that IFN-DC, according to some data, has direct cytotoxic activity [15], the use of this type of DC in chronic viral infections seems more promising.

Closest to the claimed invention is a method for immunotherapy of a viral infection, described in US Pat. hepatitis C virus (HCV) in vitro culture. The method is directed to the treatment of chronic viral infection caused, in particular, by hepatitis C, hepatitis B (HBV) and human immunodeficiency viruses (HIV). In accordance with the patent (see example No. 4 of the prototype method), DCs are generated from monocytes isolated from the fraction of mononuclear cells (MNCs) of peripheral blood of patients with HVGS in the presence of GM-CSF (1000 IU / ml) and IL-4 (800 IU / ml) for 4-5 days. The resulting immature DCs are washed and resuspended in fresh culture medium containing GM-CSF (1000 IU / ml) and IL-4 (800 IU / ml). At this stage, the cells pulsate with an antigen, which is a synthetic lipopeptide - [Th] -K (Pam2CSS) - [CTL], which consists of two virus-specific epitopes recognized by cytotoxic CD8 + lymphocytes (CTL) and T-helper cells (Th ) and interconnected by the lipid construct K (Pam2CSS), which is also a ligand for Toll-like receptors of type 2 (TLR-2). This antigen is added to immature DCs at a final concentration of 20 nM and incubated for 4 hours at 37 ° C in a humid atmosphere with 5% CO ₂ . After that, antigen-pulsed DCs are washed by centrifugation, resuspended in fresh culture medium containing GM-CSF (1000 IU / ml) and IL-4 (800 IU / ml), and cultured for 2 days. Since the antigenic construct used is simultaneously a ligand for TLR-2, it is able to induce the final differentiation of DCs without adding any additional maturing stimuli. As a result, mature DCs loaded with a synthetic lipopeptide are generated. The obtained cells are washed, resuspended in a physiological 0.9% sodium chloride solution with 10% human serum albumin (HSA) and injected intradermally into the abdomen (at a dose of 10 million cells in 1 ml) and intravenously, dropwise for 30 minutes (at a dose 10-50 million in 100 ml). If a larger number of DCs is obtained, then the remaining cells can be cryopreserved (at a dose of 20 million / ampoule) and used if necessary for re-vaccinations.

However, the prototype method has a number of significant disadvantages. So, from the description of the patent it is not clear whether this approach really allows you to induce a specific immune response against viral antigens (HCV). If so, what is the severity and duration of the immune response, and how does this affect the clinical course of chronic viral infection. Accordingly, it is not known whether this method will be effective in the treatment of herpes virus infection, which differs from other chronic viral infections caused by HCV, HBV and HIV, frequent exacerbations (up to 20 relapses per year), and that dictates the need not only to induce adequate antigen- specific immune response, but also maintaining / maintaining it at a certain level for a long period of time.

The method proposed by the authors is technologically complex and expensive. In particular, for the isolation of MNCs from peripheral blood and the subsequent production of monocytes from them, separation procedures on CliniMACS equipment (Miltenyli Biotec) are required using the appropriate disposable consumables / reagents (sets) using the Spectra MNC and CliniMACS Tubing Set / CliniMACS CD14 programs. At the same time, in order to obtain the required number of CD14 + monocytes, blood separation procedures can be repeated. In addition, in the process of generating mature, antigen-pulsed DCs, it is required to repeatedly update the culture medium with the addition of expensive recombinant cytokines GM-CSF (1000 IU / ml) and IL-4 (800 IU / ml). It is also important to note that the process of DC generation is time-consuming (up to 7 days), which significantly increases the requirements for laboratory support of sterility of cultural work.

The technological difficulties and economic costs of the prototype method are largely due to the fact that the authors propose using IL4-induced DCs (IL4-DC) for the treatment of chronic viral infections. In addition, it is proposed that a significant portion of the obtained IL4-DC be administered intravenously. However, it is known that this type of DC is characterized by low migration activity and, under conditions of growth factor deficiency, can undergo reverse transformation into monocytes. Therefore, not only the protocol for generating DCs, but also their introduction into the systemic circulation is not optimal, since such technological methods are associated with a weakening of the antigen-presenting activity of the used DCs, which can negatively affect the efficiency of the induction of the antiviral immune response in the process of DC vaccinations.

In addition, it is well known that the effectiveness of specific immunotherapy largely depends not only on the immunogenicity of the vaccine, but also on the simultaneous use of non-specific immunotherapy using various adjuvants. However, in the prototype method, this fundamental principle of active specific immunotherapy is completely ignored.

The objective of the invention is to provide a simpler and more cost-effective method of immunotherapy of a chronic often recurring herpes virus infection, which allows to reduce the number of exacerbations and the severity of their clinical manifestations.

The task is achieved by using autologous IFN-alpha-generated DCs generated from peripheral blood monocytes loaded with a recombinant herpes virus antigen in an in vitro culture, which are administered to the patient subcutaneously in 2 courses in combination with recombinant interleukin-2 as an adjuvant.

The technical result consists in the effective induction of an antigen-specific immune response and maintaining it at an adequate level for a long period of time.

The claimed method is as follows.

Hospitalization of the patient is carried out during the period of remission (at least 2 weeks after the last relapse). The indication for specific immunotherapy is the presence of a chronic herpes virus infection that is resistant to antiviral drugs and with an exacerbation frequency of more than 6 times a year.

DCs are generated from monocytes of the patient’s peripheral blood. The hemoexfusion procedure is carried out in the morning. To do this, 200 ml of the patient's venous blood are taken into a sterile vial containing a heparin solution (10,000 U), 40 ml of gelatinol solution are added and incubated for 45 min at 37 ° C. The collected leukemia suspension is collected in a separate bottle, centrifuged at 1000 rpm for 20 minutes. The autoplasma contained in the supernatant is collected for subsequent use. Precipitated leukoprecipitated cells are washed once with phosphate-buffered saline (PBS), layered on the density gradient of ficoll-verographin and centrifuged for 20 min at 3000 rpm. Mononuclear cells (MNCs) collected from interphase are washed twice and resuspended in RPMI-1640 culture medium supplemented with 5% autoplasma, 5 mM HEPES buffer and gentamicin at a dose of 80 U / ml. Then, the selected MNCs at a concentration of 3 × 10 ⁶ / ml were incubated for 2 hours at 37 ° C in sterile vials with an area of 75 or 150 cm ² (Falcon). The fraction of cells not attached to the plastic is removed. To obtain immature DCs, the complete culture medium supplemented with 1.5% autoplasm, 40 ng / ml GM-CSF (Sigma-Aldrich) and 1000 U / ml IFN-alpha (Roferon-A, Roche, Switzerland) and cultured for 3 days at 37 ° in a CO ₂ incubator at 5% CO ₂ . The final maturation of immature IFN-DK is carried out for an additional 24 hours in the presence of the pharmacopeia drug polyoxidonium (NPO PetrovaxPharm, Moscow) at a dose of 2 ng / ml. The resulting mature IFN-DK is loaded (pulsed) with a specific recombinant herpes virus antigen - HSV1gD (NPO Diagnostic Systems, Nizhny Novgorod) at a dose of 5 μg / ml during incubation at 37 ° C. Thus, the total time for obtaining IFN-DK loaded with a specific viral antigen is 4 days. Then, IFN-DK is cryopreserved in a solution of 10% DMSO and 90% albumin at a concentration of 6.0 × 10 ⁶ / ml and stored at -80 ° C until subsequent use.

Specific immunotherapy is carried out in the form of 2 courses of DC vaccinations. The first “induction” course includes 4-6 subcutaneous injections (in the upper third of the shoulder) of IFN-DK in a dose of 5 × 10 ⁶ cells with a 2-week interval. 3 months after the completion of the first course, a second “supporting” course of DC vaccinations is carried out in the form of 3-6 subcutaneous injections of IFN-DC at a dose of 5 × 10 ⁶ cells with a frequency of 1 time per month. A prerequisite for conducting DC vaccinations is the use of a recombinant human interleukin-2 human (rIL-2, Roncoleukin ^® , Biotech LLC, St. Petersburg) as an adjuvant, which is also administered subcutaneously at a dose of 0.25 mg. The use of rIL-2 as an adjuvant can significantly enhance the antigen-specific T-cell response when conducting DC vaccinations by overcoming the state of T-cell anergy, preventing T-cell apoptosis, generating T-memory cells, including long-lived cytotoxic T lymphocytes.

To assess the effectiveness of induction / maintenance of an antigen-specific immune response in patients, the level of the proliferative response of MNCs to stimulation with vaccine antigen (HSV1gD) and mitogen (concanavalin A) is examined. Studies are carried out before the start of therapy, after the completion of the 1st and 2nd courses of DC vaccination, as well as 6 months after completion of treatment (on average 15 months after the start of treatment). To do this, from 10 ml of heparinized venous blood of patients, MNCs are isolated by centrifugation in a density gradient of ficoll-verographin (as described previously). MNCs at a concentration of 0.1 × 10 ⁶ / well were cultured at 37 ° C in a humid atmosphere with 5% CO ₂ in 96-well round-bottom immunoassay plates in a complete culture medium containing RPMI-1640 supplemented with 0.3 mg / ml of L-glutamine, 5 mM HEPES buffer, 80 μg / ml gentamicin and 10% inactivated serum of AB (IY) group donors. Concanovalin A (ConA, Sigma) at a dose of 15 μg / ml and a specific recombinant herpes virus antigen - HSV1gD at a dose of 5 μg / ml are used to stimulate cells. The proliferation rate was evaluated after 72 hours by the incorporation of ³ H-thymidine into the nucleoprotein fractions, introduced 18 hours before the end of cultivation at a dose of 1 μg / ml. The radioactivity is calculated in a liquid scintillation counter SL-30 (Intertechnic, France). The results are presented as the average score (cpm) of three identical cultures.

The inventive method of immunotherapy was used in 7 patients with orofacial localization of herpes virus infection. The duration of the disease averaged 18.3 ± 1.3 years (from 15 to 25 years), and the number of exacerbations from 8 to 15 per year (six months before the start of treatment, an average of 6 ± 0.5 relapses). The treatment was carried out against the background of complete cancellation of antiviral drugs (due to their toxicity and / or low efficiency) and any other immunomodulators (interferon inducers, recombinant interferon 2-alpha, lycopid, polyoxidonium preparations, etc.). Conducting specific immunotherapy according to the developed scheme was characterized by good tolerance, did not cause local and systemic allergic and / or inflammatory reactions.

The inventive method was characterized by pronounced clinical efficacy (figure 1). A significant decrease in the number of exacerbations was recorded in patients: during treatment, the number of relapses decreased to 4.0 ± 0.7 against 6 ± 0.5 episodes before treatment (p _u <0.05), and within 6 months after the end of treatment, to 2.4 ± 0.9 (p _u <0.05). Accordingly, the inter-relapse period during therapy increased to 82 ± 14 days, and during the 6-month follow-up period after therapy was 67 ± 16 days, which was significantly higher than the corresponding indicator before treatment (18.7 ± 3.2; p _u <0, 05). Moreover, in 3 patients during the 6-month follow-up, not a single exacerbation was recorded, while in 4 patients who had relapses, exacerbations proceeded in a milder form with a smaller area of rashes and less pronounced intoxication.

A study of the antigen-specific and mitogen-induced proliferative response in cultures of MNCs of patients showed that initially patients with herpes virus infection had a reduced level of Con-A-stimulated proliferation (12314 ± 1840 versus 59240 ± 3990 imp / min in healthy donors, p _u <0, 05) and the lack of reactivity to stimulation with a viral antigen (Table 1).

After the 1st course of DC vaccination (3 months after the start of treatment), the _{hypertension AH} increased on average by 5 times, which indicated the appearance of antigenic reactivity of MNCs. However, the intensity of mitogenic reactivity of T cells remained low. At the end of the 2nd course of DC vaccination (12 months after the start of treatment), the intensity of the antigen-specific response remained at a fairly high level (IH _AH 3.8 ± 1.6 calculated units), with a 2-fold increase total mitogenic reactivity of T cells (p _U <0.05). 6 months after completion of treatment, the intensity of the antigen-specific response in patients remained at a high level and corresponded to the level of response after the first “induction” course of DC vaccination. In addition, the restoration of mitogenic T cell reactivity to a level comparable to the values of healthy donors (59240 ± 3990 imp / min) was noted.

Table 1 The dynamics of the proliferative response of MNC patients to HSV1gD antigen and ConA Survey Proliferative activity (cpm) MNC MNC + AG MNC + ConA Before treatment 198 ± 44 208 ± 44 (1.1 ± 0.23) 12314 ± 1840 (58.4 ± 8.9) After 1 IT course 167 ± 45 932 ± 280 ^∗ (5.2 ± 2.3) 16444 ± 2861 (112 ± 24.6) After 2 course IT 150 ± 27 598 ± 222 ^∗ (3.8 ± 1.6) 25979 ± 4453 ^∗ (186 ± 36) 6 months after IT 208 ± 43 1196 ± 552 ^∗ (5.4 ± 2.8) 64 450 ± 18050 ^∗ (306 ± 59)

Note: the data are presented as M ± SE of the proliferative response of MNCs in patients with herpesvirus infection in the absence and presence of AH or ConA. In parentheses is the index of the effect of IV _AG and IV _KON-A - the ratio of the stimulated response to the level of spontaneous proliferation, ^∗ - the significance of the differences compared with the pre-treatment parameters, p _u <0.05, Wilcoxon-Mann-Whitney u-test.

Thus, the immunotherapy of the claimed method was accompanied by the induction of an antigen-specific cellular immune response, which was maintained at an adequate level for at least 6 months after completion of treatment. In addition, normal proliferative reactivity of T cells in response to mitogenic stimulation was normalized in patients.

For a better understanding of the essence of the claimed invention, as well as to confirm the compliance of the solution with the condition of industrial applicability, we give examples of specific implementations with which it cannot be exhausted.

Example 1

Patient Z., 24 y., IS No. 684, was observed in the immunology department of the immunopathology clinic of the Research Institute of CI of the Siberian Branch of the Russian Academy of Medical Sciences from May 15, 2008 to September 7, 2009. Upon admission to the department, she complained of recurrent vesicular rashes on the red border of the lips with a frequency Once a month, an increase in body temperature to subfebrile values (37.2-37.3 ° C) mainly in the afternoon, increased fatigue, weakness, and decreased working capacity.

According to the anamnesis, he considers himself ill for about 17 years. Herpetic eruptions recurred more than 5 times a year, antiviral drugs were used for local treatment. Within three years before treatment, the recurrence rate increased to monthly exacerbations and amounted to 12 times / year. At the same time, a gradual increase in clinical manifestations was noted in the form of an increase in the area of the lesion, the appearance of symptoms of general intoxication, subfebrile condition both during the exacerbation period and during the relapse period. Relapses were triggered by hypothermia and / or acute respiratory infections. In treatment, she used systemic and local antiviral drugs with a short-term incomplete effect.

To generate dendritic cells 05/15/2008 hemoexfusion of peripheral blood was performed in a volume of 200 ml. After a 3-day cultivation of the adherent fraction of mononuclear cells in the presence of GM-CSF (40 ng / ml) and IFN-alpha (1000 U / ml), followed by maturation for 24 hours in the presence of polyoxidonium (2 ng / ml), the obtained dendritic cells loaded ("pulsed") with the recombinant herpes virus antigen (HSV1gD at a dose of 5 μg / ml, 1 hour at 37 ° C). The obtained antigen-specific IFN-DK was cryopreserved at -80 ° C in a solution of 10% DMSO and 90% albumin in aliquots of 5 × 10 ⁶ cells for subsequent use. The “inductor” course of DC vaccination consisted of 4 subcutaneous cell injections with an interval of 11-14 days (05.21.08; 02.06.08; 06.16.08 and 06.27.08). Dendritic cells were injected subcutaneously into the upper third of the shoulder at 4 points in the amount of 1.25 × 10 ⁶ IFN-DK at each point. Recombinant human interleukin-2 (Roncoleukin) at a dose of 0.25 mg, subcutaneously, was used as adjuvant. Against the background of the course of exacerbations was not observed.

A “supportive” immunotherapy course was started on September 10, 2008. Repeated venous blood hemoexfusion was performed with a volume of 200 ml, and IFN-DK loaded with recombinant herpes virus antigen was generated from the fraction of adhesive cells (as described earlier). Five DC vaccinations were carried out with an interval of 2-4 weeks (15.09.08; 13.10.08; 27.10.08; 10.11.08 and 24.11.08). Conducting specific immunotherapy was characterized by good tolerance, did not cause local and systemic allergic or inflammatory reactions.

Against the background of the “supportive” course of immunotherapy, two exacerbations of labial herpes were noted (after the 1st and 2nd vaccination), but the patient subjectively noted a decrease in the lesion area to 1-2 vesicles, with rapid independent regression. According to the follow-up data, the first relapse after the end of specific immunotherapy was noted in September 2009, that is, 9 months after the end of treatment. Moreover, the patient also noted a significantly smaller area of herpetic eruptions compared with the initial level.

In order to evaluate the effectiveness of induction / maintenance of an antigen-specific immune response in a patient, the level of the proliferative response of MNCs to stimulation with vaccine antigen (HSV1gD, 5 μg / ml) and mitogen (concanavalin A, 15 μg / ml) was studied. Before the start of immunotherapy (May 15, 2008), the following were recorded: low mitogenic T cell reactivity (8326 imp / min versus 59240 ± 3990 imp / min in healthy donors) and the absence of a proliferative response to the antigen (216 imp / min; IV _{arterial hypertension} = 1, 1 calculation unit). After the first and second courses of DC vaccination (September 10 and December 1, 2008, i.e., 3 and 6 months after the start of treatment), a gradual increase in the level of Con-A-stimulated proliferation to 23651 and 27500 imp / min, respectively, was noted. There was also a 1.5-2-fold increase in the antigen-specific response (IV _AG = 1.5 and 2.0 calc. Units). 6 months after completion of the maintenance course (05/12/2009, i.e., one year from the start of treatment), the mitogenic reactivity of T cells fully recovered (46,400 imp / min), and the intensity of the antigen-specific response was 796 imp / min ( IV _AG = 4.4 calculation units).

Thus, the activation of an antigen-specific antiviral immune response and the restoration of the functional reactivity of T cells, which are achieved by a course of treatment in the form of subcutaneous vaccinations of autologous IFN-alpha induced DCs loaded with the recombinant herpes virus antigen in combination with rIL-2 as an adjuvant , reduces the number of exacerbations and the severity of their clinical manifestations in patients with chronic often recurring herpesvirus infection.

Claims (1)

A method for immunotherapy of a chronic viral infection, comprising administering to a patient autologous dendritic cells generated from peripheral blood monocytes and loaded with a viral antigen in an in vitro culture, characterized in that the dendritic cells are cultured in the presence of interferon alpha, and a recombinant herpes virus antigen is used and use the obtained dendritic cells in 2 courses by subcutaneous vaccination in combination with a recombinant human interleukin-2 preparation as an adjuvant coagulant for the treatment of frequently relapsing herpes virus infection.

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