基于纳米递送系统的肿瘤免疫治疗研究进展

近年来,肿瘤免疫治疗发展迅速,相较于其他传统肿瘤治疗方法具有显著优势.肿瘤免疫治疗是通过调动或激发机体自身的免疫功能,从而抑制和杀伤肿瘤细胞.随着纳米技术的发展,生物纳米载体材料为疫苗的开发提供了全新的思路.纳米疫苗是基于纳米技术开发的治疗性或预防性疫苗,大多数肿瘤疫苗包括诱导免疫应答的外源性抗原、递送抗原的载体以及增强免疫原性并加速和延长肿瘤疫苗效果的佐剂.纳米递送载体具有良好的生物相容性和独特的理化性质,能够有效地递送各种抗原,通过调控抗原在抗原提呈细胞内的呈递途径,在激发机体的体液免疫基础上,可以进一步激活抗原特异性的细胞免疫反应.旨在对纳米递送体系在肿瘤疫苗研究中的应用作一综述.     

重组人Hsp70提呈的H22肝癌细胞混合抗原肽治疗小鼠肿瘤的研究

目的：探讨B22肝癌细胞混合抗原肽体内抑制小鼠肿瘤的最佳方式。方法：采用冻融，低渗振荡，加热沉淀及酸处理等方法从H22肝癌细胞中制备抗原肽；将接种过H22瘤细胞的小鼠分别给予Hsp70-H22肽复合物注射，复合物＋pCH510质粒注射或化疗后给予复合物＋质粒注射，观察肿瘤抑制情况。结果：上述方法制备的抗原肽为混合肽，其与Hsp70形成的复合物体内能够抑制小鼠肿瘤生长，并且这种复合物可与pCH510质粒协同完全抑制10^5接种的瘤细胞形成肿 瘤，但不能完全抑制10^6接种的瘤细胞形成肿瘤，然而这种双因素与化疗进行衔接则可完全抑制10^6接种的瘤细胞形成肿瘤。结论：H22肝癌细胞肿瘤混合抗原肽经rhHsp70提呈可诱导H22荷瘤小鼠产生特异性免疫保护作用，与其它因素联合应用可产生更好的治疗效果。     

树突状细胞亚群及其在疾病中的作用研究进展北大核心CSCD

树突状细胞(DCs)来源于骨髓造血干细胞(HSCs)的一类高度异质性细胞,作为体内极其重要的专职性抗原提呈细胞和免疫协调器,具有显著的细胞表型和功能可塑性。近年来,通过高通量分析等技术发现了一些新的DCs亚群。免疫微环境赋予了DCs不同的亚群特征,不同的DCs亚群在不同的组织器官或疾病中发挥免疫调节作用。并为肿瘤移植排斥、自身免疫性疾病等疾病的治疗提供新思路。本文综述了DCs亚群的研究进展以及其在免疫相关性疾病中的作用。     

免疫抑制剂作为DNA疫苗佐剂的研究

目的:探讨免疫抑制剂作为DNA疫苗佐剂的免疫效果及预防自主免疫疾病的效果.方法:将三种免疫抑制剂环孢素(C&A)、他克莫斯(FK506)和霉酚酸酯(MMF)作为多发性硬化症(MS)的DNA疫苗佐剂,共同肌肉注射免疫小鼠.取小鼠脾脏抗体染色后,用流式细胞仪检测调节性T细胞(Treg)的比例及T细胞增殖反应,检测树突细胞(DCs)成熟状态及分泌白介素10(IL-10)的变化,并在诱导EAE动物模型后,检测临床打分的变化.结果:免疫FK506作为MS的DNA疫苗佐剂组显著提高了Treg细胞的比例,T细胞增殖反应显著受到了抑制,DCs细胞成熟受到抑制且IL-10的表达显著升高,能够显著降低小鼠实验性变态反应性脑脊髓炎(EAE)的临床打分.结论:FKS06能够作为DNA疫苗的佐剂,诱导Treg细胞的产生,使机体产生免疫耐受,并能够在一定程度上预防自主免疫疾病的发生,为自身免疫性疾病的预防和治疗提供新的方法和思路.     

Hsp70-肿瘤抗原肽复合物修饰的DC疫苗体内外特异性抗瘤作用

目的 :探讨树突状细胞 (DC)及Hsp70 肿瘤抗原肽复合物修饰后的体内外特异性抗瘤作用。方法 :采用一定的生化技术 ,从H2 2肝癌细胞中提取肿瘤抗原肽 ,并在体外与Hsp70进行结合 ;采用细胞培养技术 ,培养rmGM CSF、rmIL 4诱导的小鼠骨髓细胞 ,体外获取大量的DC ,后者经Hsp70 肿瘤抗原肽复合物修饰后刺激小鼠脾淋巴细胞 ,通过MTT法进行检测淋巴细胞的激活 ;收集上述刺激传代培养的淋巴细胞 ,检测其对H2 2瘤细胞和艾氏腹水癌细胞的杀伤功能 ;采用H2 2瘤细胞肌肉接种和H2 2瘤细胞、艾氏腹水癌细胞腹腔接种 ,对接种小鼠给予经体外修饰的DC回输 ,观察其抑制肿瘤的效果。结果 :Hsp70 肿瘤抗原肽复合物可使DC成熟 ,大量分泌IL 12、TNF α、IL 1β等细胞因子 ,并能够使DC激活小鼠脾淋巴细胞 ;激活后传代培养的淋巴细胞对H2 2瘤细胞能够特异性地杀伤 ,而对艾氏腹水癌细胞无效 ;经Hsp 70 肿瘤抗原肽复合物修饰后的DC可作为一种有效的瘤苗 ,体内能特异性地抑制小鼠H2 2肿瘤生长。结论 :Hsp70 肿瘤抗原肽复合物能够很好地修饰体外诱导获取的DC ,使后者成为一种有效的瘤苗 ,体外能够特异性地激活淋巴细胞 ,体内有效地抑制肿瘤生长     

非细胞因子来源的天然蛋白质作为人类疫苗佐剂的研究进展

疫苗接种不仅是现代医学预防传染病的常用方式和重要手段,也已成为肿瘤预防与治疗的研究热点。疫苗效能的充分发挥离不开佐剂的应用。许多疫苗本身含有非特异性佐剂,它们可以增强疫苗刺激机体产生免疫应答的作用。研究发现,多种生物来源的天然蛋白质也可作为佐剂应用于疫苗接种中,并且与传统佐剂相比,天然蛋白质用作佐剂有其独特的优势。本文主要就非细胞因子来源的天然蛋白质作为疫苗佐剂的应用现状及其免疫机制的最新研究进展进行综述。     

CpG-ODN疫苗佐剂的研究进展

含胞嘧啶-鸟嘌呤（ CpG ）二核苷酸的寡聚脱氧核苷酸链（ CpG-ODN）是一种具有巨大开发潜力的佐剂，这是一类合成的、包含有1个或多个非甲基化的CpG基序的单链DNA。研究表明， CpG-ODN能够选择性地增强脊椎动物的细胞和／或体液免疫应答，有效提升多种疫苗的免疫效果［1］。它能直接活化树突状细胞、巨噬细胞和B细胞，增强抗原提呈能力，促进抗体的分泌；间接活化T细胞和NK细胞等免疫效应细胞，增强其功能及细胞因子的分泌，诱导Th1型免疫应答，产生细胞及体液免疫，因此， CpG-ODN是一种理想的疫苗佐剂。目前国外研究CpG-ODN作为感染性疾病和肿瘤疫苗佐剂已进入Ⅰ／Ⅱ期临床试验阶段［2，3］，并已证实CpG-ODN是一种高效低毒的新型免疫佐剂。     

黄芩素作为佐剂对抗原提呈细胞的免疫增强作用

目的从体外和体内2个方面检测黄芩素对抗原提呈细胞的免疫增强活性,为促进临床肿瘤免疫治疗的发展提供实验基础。方法在体外实验中,以LPS作为阳性对照,不同浓度的黄芩素分别刺激DC2.4细胞系和RAW264.7细胞系,应用流式细胞仪检测2种细胞表面CD80、CD86、MHC-Ⅰ、MHC-Ⅱ表达量变化;在体内试验中,以弗式完全佐剂与灭活的肿瘤抗原作为阳性对照,黄芩素与灭活的肿瘤抗原作为实验组对小鼠进行免疫,免疫3 d后对其免疫效果进行评价。结果在体外实验中,不同浓度的黄芩素分别刺激DC2.4和RAW264.7细胞后,细胞表面CD80、CD86、MHC-Ⅰ、MHC-Ⅱ都有不同程度表达上调,在体内实验中,不同浓度的黄芩素可使抗原提呈细胞表面CD80、CD86、MHC-Ⅰ、MHC-Ⅱ都有不同程度表达上调,同时使细胞因子IL-12和TNF-α有不同程度表达上调,而使IL-10表达下调。结论黄芩素在体外和体内具有较强的激活抗原提呈细胞的作用,并且使抗原提呈细胞表面标志和细胞因子上调,黄芩素有可能成为具有潜在应用价值的肿瘤细胞疫苗佐剂。     

树突状细胞直接抗肿瘤作用研究进展

树突状细胞(DC)是功能最强的抗原递呈细胞,其除可将肿瘤抗原递呈给T细胞以间接杀伤肿瘤细胞外,还可作为效应细胞直接发挥抗肿瘤作用.DC可通过细胞间接触和分泌细胞因子直接抑制肿瘤细胞的增殖,并可通过与TNF家族密切相关的凋亡通路直接杀伤敏感肿瘤细胞.DC的这一固有免疫功能在肿瘤免疫治疗中具有重要的意义.     

免疫脂质体与临床

将特异性抗体粘附于脂质体上形成免疫脂质体,它能特异性识别表达具有某种抗原的细胞或组织.由它携带药物具有靶向性强,毒副作用小,半衰期长等优点.针对其特点,临床应用免疫脂质体开展了对多种疾病的治疗.根据免疫脂质体携带药物对疾病治疗的优缺点,本文综述了近几年的临床观察和研究的进展.     

Differential changes in heat shock protein-, lipoarabinomannan-, and purified protein derivative-specific immunoglobulin G1 and G2 isotype responses during bovine Mycobacterium avium subsp. paratuberculosis infection

Bovine paratuberculosis is caused by infection of young calves with Mycobacterium avium subsp. paratuberculosis. In some of the chronically infected cows the long asymptomatic stage (2 to 4 years) is followed by a rapid progression to a clinical stage due to protein-losing enteropathy, which will ultimately be fatal. The current dogma is that in early stages of disease the cell-mediated responses predominate, whereas in the clinical stage of the disease the humoral responses prevail, possibly signaling a switch in immune reactivity related to disease progression. We developed immunoglobulin M (IgM)-, IgA-, and IgG1- and IgG2-isotype-specific enzyme-linked immunosorbent assays for M. avium subsp. paratuberculosis-derived antigens (heat shock proteins of 70 kDa [Hsp70] and 65 kDa [Hsp65], lipoarabinomannan [LAM], and M. avium subsp. paratuberculosis purified protein derivative PPD [PPDP]). The serological responses of cows in different stages of paratuberculosis were used to evaluate the putative shift in immune responsiveness. In the clinical stage the PPDP-specific IgG1 responses were increased compared to those in the asymptomatic stage. However, total IgG1 and IgG2 and the Hsp70-, Hsp65-, and LAM-specific isotype responses were decreased in the clinical stage were decreased compared to those in the asymptomatic stage of disease. Thus, the classical pattern was found only for PPDP antigens and the IgG1 isotype. For other antigens and isotypes and the total IgG levels, the response pattern is different and indicates that there is no uniform association with increased antibody responses during the progression from the asymptomatic stage to the clinical stage of bovine paratuberculosis.     

Murine peritoneal macrophages activated by the mycobacterial 65-kilodalton heat shock protein express enhanced microbicidal activity in vitro.

After an intraperitoneal (i.p.) injection of purified protein derivative, peritoneal macrophages from mice infected with Mycobacterium bovis bacillus Calmette-Guérin (BCG) show an enhanced respiratory burst, inhibit the intracellular proliferation of Toxoplasma gondii, and kill Listeria monocytogenes more efficiently than peritoneal macrophages from normal mice. One of the immunodominant antigens of Mycobacterium spp. is the 65-kDa heat shock protein (Hsp 65), and in the present study, we determined whether injection of this protein into mice leads to activation of their peritoneal macrophages. After an i.p. injection of Hsp 65, peritoneal macrophages from BCG-infected CBA/J mice also released more H2O2, inhibited the proliferation of T. gondii, and killed L. monocytogenes faster than peritoneal macrophages from normal mice, although Hsp 65 was less effective than purified protein derivative. When normal mice were injected with Hsp 65 suspended in saline after a booster injection with Hsp 65, their macrophages did not display enhanced antimicrobial activity, indicating that an adjuvant was required for a cellular immune response against Hsp 65. In the present study, the adjuvant dimethyl dioctadecylammonium bromide (DDA) was preferred because it contains no endotoxin or mycobacterial antigens and because it has been reported that DDA does not induce the production of gamma interferon. Peritoneal macrophages from C57BL/6 and CBA/J mice that had received a subcutaneous injection of Hsp 65 suspended in DDA followed by an i.p. booster injection of Hsp 65 suspended in saline were activated, as indicated by the enhanced production of H2O2, inhibition of the intracellular proliferation of T. gondii, and increased rate of intracellular killing of L. monocytogenes in vitro relative to that by resident peritoneal macrophages and peritoneal macrophages obtained from mice that had received ovalbumin instead of Hsp 65. The rate of phagocytosis of L. monocytogenes was not affected by Hsp 65 treatment. Despite the in vitro expression of enhanced microbicidal activity of peritoneal macrophages, no difference in the growth of L. monocytogenes in the liver and spleen between Hsp 65-treated and control mice was found.     

Over-expression of gene encoding heat shock protein 70 from Mycobacterium tuberculosis and its evaluation as vaccine adjuvant

Background: Heat shock proteins (Hsps) are evolutionary ancient and highly conserved molecular chaperons found in prokaryotes as well as eukaryotes. Hsp70 is a predominant member of Hsp family. Microbial Hsp70s (mHsp70s) have acquired special significance in immunity since they have been shown to be potent activators of the innate immune system and generate specific immune responses against tumours and infectious agents. Objectives: The present study was aimed to clone express and purify recombinant Hsp70 from the Mycobacterium tuberculosis and characterise it immunologically. The study also aimed at determining the potential of recombinant M. tuberculosis heat shock protein (rMTB-Hsp70) as adjuvant or antigen carrier. Materials and Methods: Cloning of M. tuberculosis heat shock protein (MTB-Hsp70) amplicon was carried out using the pGEMT-Easy vector although for expression, pProExHTb prokaryotic expression vector was used. Purification of recombinant Hsp70 was carried out by nickel-nitrilotriacetic acid (Ni-NTA) affinity chromatography. For immunological characterization and determining the adjuvant effect of MTB-Hsp70, BALB/c mice were used. The data obtained was statistically analysed. Results: Hsp70 gene was cloned, sequenced and the sequence data were submitted to National Center for Biotechnology Information (NCBI). Recombinant MTB-Hsp70 was successfully over-expressed using the prokaryotic expression system and purified to homogeneity. The protein was found to be immunodominant. Significant adjuvant effect was produced by the rMTB-Hsp70 when inoculated with recombinant outer membrane protein 31; however, effect was less than the conventionally used the Freund’s adjuvant. Conclusion: Protocol standardised can be followed for bulk production of rHsp70 in a cost-effective manner. Significant adjuvant effect was produced by rMTB-Hsp70; however, the effect was than Freund’s adjuvant. Further, studies need to be carried out to explore its applicability as carrier of antigen.     

Mycobacterial Heat-Shock Protein 65 Induces Proinflammatory Cytokines but does not Activate Human Mononuclear Phagocytes

The 65 kDa heat-shock protein (Hsp65), a well-conserved and immunodominant antigen which elicits a cellular and humoral immune response, may play a role in host defence against invading microorganisms and autoimmune disorders. The aim of the present study was to assess the effects of Hsp65 on the functional activities of human mononuclear phagocytes in the absence of lymphocytes. Incubation with Hsp65 resulted in an enhanced release of TNF-γ and IL-1γ by human monocytes and monocytederived macrophages (MDM). The amount of cytokines released by these cells in response to Hsp65 was similar to that released in response to IFN-γ together with LPS. Incubation with ovalbumin did not stimulate the release of these cytokines. In vitro stimulation of monocytes with Hsp65 enhanced the membrane expression of complement receptor III but did not influence either the expression of Fc-receptor I and HLA class-II antigens or the release of reactive oxygen intermediates. Therefore, Hsp65 stimulated monocytes cannot be considered to be activated according to classical criteria. The release of the proinflammatory cytokines TNF-γ and IL-1γ by human mononuclear phagocytes in response to Hsp65 indicates that this protein can contribute to both host defence and tissue damage in inflammatory lesions characterized by an abundant expression of Hsp65.     

Initiation of adaptive immune responses by transcutaneous immunization

The development of new, effective, easy-to-use and lower-cost vaccination approaches for the combat against malignant and infectious diseases is a pre-eminent need: cancer is a leading cause of morbidity in the Western World; there are numerous pathogenic diseases for which we still have no protective or therapeutic cure; and the financial limitations of developing countries to fight these diseases. In this mini-review we focus on transcutaneous immunization (TCI), a relatively new route for antigen delivery. TCI protocols appear to be particularly promising by gaining access to skin resident APC, which are highly efficient for the initiation of humoral and/or cellular immune responses. Consisting of an adjuvant as a stimulus in combination with an antigen which defines the target, TCI offers a most attractive immunization strategy to mount highly specific full-blown adaptive immune responses. As a topically applicable cell-free adjuvant/antigen mixture, TCI might be suitable to improve patient compliance, as well as feasible economically for the use in Third World countries. In addition, this non-invasive procedure might increase the safety of vaccinations by eliminating the risk of infections related to the recycling and improper disposal of needles. The dissection of antigen and adjuvant is important because it allows "free" combinations in contrast to classical immunizations which are based on application of the pathogen of interest. The most relevant ways and means to find new, effective pathogenic target antigens are "reverse vaccinology" and the direct peptide-epitope identification from MHC molecules with mass-spectrometry. Due to these efficient approaches the variety of antigenic epitopes for potential protective/therapeutic use is perpetually expanding. The most studied adjuvants in TCI approaches are cholera toxin (CT) and its less toxic relative, the heat-labile enterotoxin (LT). Both CT and LT can serve as antigen as well. In contrast to these large proteins, which can only penetrate "pre-treated" skin barrier, the immune response modifier, TLR7 agonist R-837 (Imiquimod) is a small compound adjuvant that easily passages non-disrupted epidermis. It remains currently elusive which cells of the complex-structured "skin-associated lymphoid tissue" (SALT) respond to the adjuvant and which APC carries the antigen to the draining lymphnodes for subsequent initiation of adaptive immune responses.     

Cell-Mediated Immune Responses and Protective Efficacy against Infection with Mycobacterium tuberculosis Induced by Hsp65 and hIL-2 Fusion Protein in Mice

Heat shock protein 65 (Hsp65) is an important immunodominant antigen against tuberculosis (TB), and interleukin-2 (IL-2) plays an important role in the regulation of antimycobacteria immune responses. In order to further increase the immunogenicity of Hsp65 against infection caused by Mycobacterium tuberculosis (MTB), we expressed MTB Hsp65 and human IL-2 fusion protein, Hsp65-hIL-2, in Escherichia coli. The expression of Hsp65-hIL-2 was confirmed by Western blotting using anti-Hsp65 MoAb and anti-hIL-2 MoAb, respectively. Hsp65-IL-2 and Hsp65 were then purified by Ni-NTA affinity chromatography. Mice were immunized with purified Hsp65-hIL-2 or Hsp65 emulsified in the adjuvant combination dimethyl dioctadecylammonium bromide and monophosphoryl lipid A. Eight weeks after immunization, there was significant proliferation of spleen lymphocytes in response to both Hsp65 and Hsp65-hIL-2 proteins. Interestingly, Hsp65-hIL-2 fusion protein elicited significantly higher levels of IFN-γ and IL-2 in the lymphocytes culture supernatant than that of the BCG (Denmark strain) immunized group and Hsp65 group ( P  < 0.05). After challenging the immunized mice with MTB, the bacteria loads in the spleens and lungs of mice immunized with the fusion protein were significantly lower than Hsp65 alone group, reaching an equivalent level as BCG immunization group. Our results suggest that the Hsp65 and hIL-2 fusion protein may serve as an alternative vaccine candidate against MTB infection.     

Modulation of pristane-induced arthritis by mycobacterial antigens.

Several prominent mycobacterial protein antigens involved in antibody and T cell responses have been identified as members of highly conserved heat shock protein families. In particular, immune responses to the mycobacterial 65 kD heat shock protein (hsp65) have been implicated in the pathogenesis of autoimmune diseases both in experimental animal models and in man. Additionally, hsp65 has been shown to modulate the course of autoimmune disease in such experimental animal systems. In this report, we have examined the synthesis of heat shock proteins by a fast growing mycobacterial strain, M. vaccae, in heat stressed cultures and used the pristane induced arthritis model to investigate the immunoprophylactic and immunotherapeutic potential of heat killed M. vaccae. Heat shock of M. vaccae cultures at 48 degrees C demonstrated a 43-fold increase in hsp65 over that expressed at 37 degrees C. It is therefore suggested that heat killed M. vaccae contains sufficient hsp that can be presented in the context of appropriate adjuvant properties for use as an effective immunomodulatory agent. Immunisation experiments with M. vaccae revealed that protection or exacerbation of pristane induced arthritis was dependent on the dose (given in an oil or aqueous suspension), route and time of immunisation. In addition, it was demonstrated that the development of arthritis correlated with high levels of agalactosyl IgG and that "protected" animals had significantly depressed levels.     

A heterologous DNA priming-Mycobacterium bovis BCG boosting immunization strategy using mycobacterial Hsp70, Hsp65, and Apa antigens improves protection against tuberculosis in mice

Tuberculosis is responsible for >2 million deaths a year, and the number of new cases is rising worldwide. DNA vaccination combined with Mycobacterium bovis bacillus Calmette Guerin (BCG) represents a potential strategy for prevention of this disease. Here, we used a heterologous prime-boost immunization approach using a combination of DNA plasmids and BCG in order to improve the efficacy of vaccination against Mycobacterium tuberculosis infection in mice. As model antigens, we selected the M. tuberculosis Apa (for alanine-proline-rich antigen) and the immunodominant Hsp65 and Hsp70 mycobacterial antigens combined with BCG. We demonstrated that animals injected with a combination of DNA vectors expressing these antigens, when boosted with BCG, showed increased specific antimycobacterial immune responses compared to animals vaccinated with BCG alone. More importantly, the protection achieved with this regimen was also significantly better than with BCG alone.     

Mucosal adjuvants and anti-infection and anti-immunopathology vaccines based on cholera toxin, cholera toxin B subunit and CpG DNA

Mucosal immunisation may be used both to protect the mucosal surfaces against infections and as a means for immunological treatment of peripheral immunopathological disorders through the induction of systemic antigen-specific tolerance ('oral tolerance'). The development of mucosal vaccines, whether for prevention of infectious diseases or for oral tolerance immunotherapy, requires efficient antigen delivery and adjuvant systems that can help to present the appropriate vaccine or immunotherapy antigens to the mucosal immune system. The most potent (but also toxic) mucosal adjuvants are cholera toxin (CT) and the closely related Escherichia coli heat-labile enterotoxin (LT), and much effort and significant progress have been made recently to generate toxicologically acceptable derivatives of these toxins with retained adjuvant activity. Among these are the non-toxic, recombinantly produced cholera toxin B-subunit (CTB). CTB is a specific protective antigen component of a widely registered oral cholera vaccine as well as a promising vector for either giving rise to mucosal anti-infective immunity or for inducing peripheral anti-inflammatory tolerance to chemically or genetically linked foreign antigens administered mucosally. CT and CTB have also recently been used as combined vectors and adjuvants for markedly promoting ex vivo dendritic cell (DC) vaccination with different antigens and also steering the immune response to the in vivo-reinfused DCs towards either broad Th1 + Th2 + CTL immunity (CT) or Th2 or tolerance (CTB). Another type of mucosal adjuvants is represented by bacterial DNA or synthetic oligodeoxynucleotides containing CpG-motifs, which especially when linked to CTB have been found to effectively stimulate both innate and adaptive mucosal immune responses. The properties and clinical potential of these different classes of adjuvants are being discussed.     

Regulation of the Physiological Functions of Human Dendritic Cells by Recombinant Heat Shock Protein Hsp70

Dendritic cells (DC) are the most important antigen-presenting cells in the body. They are the target of action of various vaccines and dendritic cells have been used as the basis for developing cellular antitumor and antiviral vaccines, i.e., DC vaccines. At the same time, dendritic cells may provide a suitable model for studies of the activity and mechanisms of action of different immunotherapeutic formulations. One aspect of the optimization of the use of dendritic cells for inducing antigen-specific immune responses relates to the use of heat shock proteins (Hsp), particularly Hsp70. This protein can be used to introduce protein antigens into dendritic cells and to control the activity of dendritic cells. Important aspects of achieving these aims include knowledge of dendritic cell physiology and the characteristics of the interaction of Hsp70 and its complexes with antigens with dendritic cells of different levels of differentiation. Human recombinant Hsp70 was found not only to deliver antigens to dendritic cells, but also to regulate the activity of mature dendritic cells and to optimize the induction of antigen-specific cellular immune responses.