The importance of alpha/beta (alpha/13) interferon receptors and signaling pathways for the treatment of cervical intraepithelial neoplasias.

INTRODUCTION: Immunotherapies have been effective in treating various forms of cancer, including cervical intraepithelial neoplasias (CINs) predominantly caused by human papilloma virus (HPV). DEVELOPMENT: To establish persistent infections in stratified epithelia, HPV induces proliferative lesions. Viral gene products are able to change gene expression and cellular proteins. Interferons (IFNs) are inducible glycoproteins that have immunomodulatory, antiviral, antiproliferative, and antiangiogenic effects. In particular, interferon-alpha (IFN-alpha) has been shown to inhibit the development and progression of cervical cancer. In this review, actions of interferons alpha/beta (alpha/beta), including their receptors and signaling pathways, are described, as well as their clinical importance in the immune response against cervical lesions. CONCLUSION: The interaction of IFN-alpha/beta with its receptor results in a series of phosphorylation events. These mechanisms can be ineffective in IFN response, then it can also compromise the therapeutic effects of immunotherapy.

An evaluation of immune system cell infiltrate in the cervical stroma of patients with grade III cervical intraepithelial neoplasia after treatment with intralesional alpha-2B interferon.

The aim of this study was to characterize infiltrating immune cells in cervical stroma biopsy samples from patients diagnosed with cervical intraepithelial neoplasias (CINs) who were treated with IFN-alpha 2b. The authors studied 13 volunteers who were diagnosed with Cervical intraepithelial neoplasia CIN II or III and who received intra-lesional treatment with IFN-alpha 2b. They collected pre- and post-treatment biopsies from each patient. They also examined the slides under a common optical microscope with a X400 lens for biopsy sample sections that were labeled with immunohistochemistry for T lymphocyte, B lymphocyte, natural killer cell, macrophage, iNOS, and perforin markers. The presence of immune response cells in the lesion was observed after treatment with intralesional IFN-alpha 2b in patients with CIN II/III changes, a reduction in CD4+ and CD8+ T lymphocyte infiltration in the women who responded well to treatment. However, there was a significant increase in these markers in samples from women who did not respond to treatment. Nonetheless, immunotherapy with IFN-alpha 2b administered intralesionally in patients with CIN II/III yields favorable results in patients who do not smoke.

IL-17 and IL-22 serum cytokine levels in patients with squamous intraepithelial lesion and invasive cervical carcinoma.

PURPOSE: Several works correlate the synthesis of IL-17 and IL-22 with tumoral progression. However, there are no studies of these cytokines on cervical cancer. The authors studied the concentration of IL-17 and IL-22 on serum obtained from patients with different grades of squamous intraepithelial lesions (SILs) and invasive cervical carcinoma. MATERIALS AND METHODS: Eighty-one women were enrolled in this study, including 23 in the healthy control (with no history of infection or lesions), 11 with low-grade squamous intraepithelial lesion (LSIL), 36 with high-grade squamous intraepithelial lesion (HSIL), and 11 who were diagnosed anatomo-pathologically with invasive carcinoma. Levels of the IL-17 and IL-22 cytokines were measured in the serum obtained from these patients using the enzyme-linked immunoabsorbent assay (ELISA) method. RESULTS: IL-17 and IL-22 displayed a similar pattern of results, with an increase in the serum level of LSIL patients, when compared with serum from HSIL patients (respectively, mean- pg/ml: 22.50 vs 12.20, and 168.2 vs 61.48, p < 0.05). CONCLUSION: Concentrations of IL-17 and IL-22 in the peripheral blood of patients with LSIL were increased compared to HSIL patients.

Immune response and immunotherapy in intraepithelial and invasive lesions of the uterine cervix.

Infection with the human papillomavirus virus (HPV) induces innate and acquired immune responses in the cervical stroma, which are a delicate, balanced and generally unpredictable immunological defense. Because of the immunological breaks that the HPV virus causes, eradication of infected cells does not occur, potentially leading to development of intraepithelial and invasive lesions. Advances in our understanding of the immune system and in the definition of antigens in tumor cells has led to many new treatment strategies. As a result, immunotherapy has the potential to be the most specific treatment for tumors, and one that requires elaboration. Recently, immunotherapy with interferon and dendritc cells has been used on intrapepithelial and invasive cervical lesions with promising results.

Immunotherapy with dendritic cells for gynecological neoplasias: a new therapeutic approach?

The immune system consists of a complex collection of mediators and cells that act in a coordinated way to eliminate neoplastic cells. One of immunotherapy's promises is the development of cellular vaccines, or more specifically, vaccines with dendritic cells. However, we still have a lot left to study and learn, since we already know that patients with tumors of the same histological grade can have completely different behaviors when given the same immunological stimulus. We believe that antitumor immunotherapy will lead to a personalized vaccine, since the scheme of treatment, the stimuli and the dosages need to be tailored to each patient.

Immunological evaluation of vaginal secretion in patients with high-grade cervical intraepithelial neoplasia treated with intralesional interferon alpha-2b.

INTRODUCTION: Conservative treatment with intralesional interferon (IFN) is a therapeutic option for cervical intraepithelial neoplasia (CIN) patients of childbearing age. MATERIALS AND METHODS: The study group was made up of patients diagnosed with a high-grade lesion and treated with intralesional human recombinant IFNalpha-2b. Vaginal secretion was collected during IFNalpha-2b treatment for analysis of cytokines and viral load. RESULTS: The initial histology diagnostic was 62.5% (n = 5) with CIN 2 and 37.5% (n = 3) with CIN 3. In terms of clinical evaluation and anatomopathology, 6.5% (n = 5) had a good clinical response, while 37.5% (n = 3) had therapeutic failure. All the patients with therapeutic failure were smokers. Interleukin 6 and tumor necrosis factor-alpha concentrations were raised at the sixth application for the patient group who failed to respond to therapy compared to the responsive group (p = 0.0357). Patients with a good response exhibited a reduction in human papillomavirus viral load (p = 0.03). CONCLUSIONS: Patients that had a good response had lower concentrations of inflammatory cytokines than did non-responders.

Expression of cytokines in cervical stroma in patients with high-grade cervical intraepithelial neoplasia after treatment with intralesional interferon alpha-2b.

Conservative treatment with interferons (IFNs) has the advantage of preserving reproductive capacity in patients with grade 2 or 3 cervical intraepithelial neoplasia (CIN). The objective of this work was to study patients with high-grade CIN treated with intralesional IFN alpha-2b and to analyze the expression of Th1, Th2 and Th3 cytokines in cervical stroma. We observed that patients with a satisfactory response (60%) to treatment with IFN alpha-2b expressed more Th1 (IFN-gamma TNF-alpha, IL-2) cytokines, with a significant reduction in the viral load of high-risk human papillomavirus (p = 0.0313). All patients with therapeutic failure were smokers and had higher expression of cytokines Th2 (IL-4) or Th3 (TGF-beta2 and TGF-beta3).

The clinical use of type 1 interferon in gynecology.

Interferons were initially described in terms of their role in blocking viral replication. They are important cytokines that act on various aspects of cell physiology. Importantly, they can affect cell proliferation or induce the differentiation of neoplastic cells. The exact way in which the interferon complex (IFN) acts on tumours is still unknown, although their use in clinical practice has been widely recommended, especially with tumours that are resistant to conventional treatments, or in situations where surgical removal might lead to a loss of organ function. IFN can be employed as a treatment for various chronic diseases, including tumours. Indeed interferon cytokines are the therapy of choice in certain situations. However, one of the difficulties yet to be overcome is the need for frequent administrations of the drug. We believe that the development of new formulations is needed to match the demand for its use in oncology treatment.

Pregnancy outcome after treatment of cervical intraepithelial neoplasia by the loop electrosurgical excision procedure and cold knife conization.

PURPOSE OF INVESTIGATION: The aim of this study was to evaluate the effect of LEEP and cold-knife conization on the outcome of subsequent pregnancy in a tertiary public hospital. METHODS: One hundred and ninety-nine patients met the inclusion criteria (age between 18 and 45 years old). Cold-knife conization, LEEP, and both (conization and LEEP) were performed in 102 (51.3%), 95 (47.7%) and two (1%) women, respectively. Average ages were respectively, 33 +/- 7.3; 25 +/- 6.73 and 30 +/- 2.8. RESULTS: Pregnancies occurred 2.6 and 4.8 years after LEEP and conization, respectively. Miscarriages and preterm pregnancies were more frequent in conization cases versus LEEP, 26% and 5.2%, 23% and 5.5%, respectively. CONCLUSION: If patients express a desire for pregnancy, LEEP should be the procedure of choice.

Peri- and intratumoral T and B lymphocytic infiltration in breast cancer.

PURPOSE: To investigate peritumoral and intratumoral infiltrates in surgical specimens obtained from patients with invasive breast cancer, and of relating these to tumor size. METHODS: Twenty-six surgical specimens obtained from patients diagnosed with breast cancer underwent immunohistochemical preparation and CD3, CD8, CD20 and CD68 labeling. The positive cells were counted in the tissue samples and correlated with the tumor size determined by imaging methods (TIA < or = 2 or TIB > 2 cm). RESULTS: There was a significant reduction in intratumoral B lymphocytes (CD20+), although this reduction could only be observed in TIA. In relation to peritumoral T lymphocytes (CD3+), there was a significant reduction in TIB, in comparison with TIA. Peritumoral and intratumoral CD3+ and CD68+ presence in completely opposite ways in both sizes of tumors. CONCLUSION: Peritumoral and intratumoral infiltrates of T and B lymphocytes are different and depend on tumor size.

Frequency of infectious agents for vaginitis in patients with a cytological diagnosis of atypical squamous cells of undetermined significance.

AIM: To evaluate the presence of infectious agents for vaginitis in patients with ASCUS. METHODS: 33,388 patients who underwent cervical-vaginal cytology from 08/1993 to 05/2002 were included in the study, and 1,104 (3.4%) presented positive ASCUS. The appraised infectious agents were Coccobacilli, Candida sp, Trichomonas vaginalis, and clue cells (Gardnerella vaginalis). RESULTS: In the group with ASCUS a larger frequency of Coccobacilli (22.37%) and Trichomonas vaginalis (5.25%) was found when compared with the group with negative ASCUS (17.79% and 3.98%, respectively; p < 0.05). Cytolysis occurred more frequently in patients with ASCUS (3.8 vs 6.3%, first phase and 4.5 vs 10%, second phase). CONCLUSIONS: We believe that some diagnoses of ASCUS can be induced by the presence of infectious agents for vaginitis, mainly cocci and coccoides. ASCUS occurs more frequently in the first phase of the menstrual cycle, therefore in less acid vaginal pH.

Potential therapeutic vaccine strategies and relevance of the immune system in uterine cervical cancer.

The interaction of HPV with the immune system has been studied, but the results are still inconclusive for several reasons. Until now, we have not been able to understand the mechanisms of immune system regulation in the uterine cervix. HPV infection does not unleash an inflammatory response, and consequently an efficient and specific immune response against the virus. Moreover, an understanding of HPV infection and local immune response is indispensable for the development of new bioactive drugs and therapies for patients with both non invasive and invasive tumors, mainly for patients that do not present regression with radiotherapy or chemotherapy or in whom the tumors are surgically unresectable. The aim of this review is to provide support in understanding potential mechanisms used by the immune system to destroy neoplastic cells, comparing the immunotherapy used in cancer and discussing the possibility of developing new drugs based on these mechanisms of action.

T lymphocytes (CD3) may participate in the recurrence of cervical intraepithelial neoplasia grade III.

OBJECTIVE: Data from the literature demonstrate that the local and systemic immune responses seem to play an important role in the progression of cervical intraepithelial neoplasia (CIN). Our aim was to investigate whether recurrences among CIN III patients might be related to the presence of local lymphocytes, macrophage and enzyme iNOS. METHODS: We analyzed 35 patients with CIN III who underwent conization and followed up for a minimum of 4 years. Using immunohistochemistry, the presence of T lymphocytes (CD3, CD8 and CD45RO), B lymphocytes (CD20), macrophages (CD68) and the expression of the enzyme iNOS were investigated. The quantity of marked cells is graded as: 0, absence of cells; 1, rare cells; 2, moderate number of cells; 3, many cells. For statistical purposes, we took the scores 0 and 1 to indicate weak marking and the scores 2 and 3 to indicate strong marking. RESULTS: We found strong positive expression of CD3-positive T lymphocytes among CIN III patients with recurrence following conization (100 vs. 50% without recurrence, p=0.02). We did not find any statistical differences in the expression of CD20, CD68, CD45RO, CD8 or iNOS. CONCLUSIONS: It is concluded that strong positive findings of CD3 T lymphocytes were related to recurrence following conization due to CIN III.

Trypanosoma cruzi: the role of PGE2 in immune response during the acute phase of experimental infection.

Chagas disease is characterized by cardiac lesions and a high level of PGE2. Our objective was to investigate the role of PGE2 in cardiac lesions. BALB/c mice were infected with Trypanosoma cruzi (1x10(3) trypomastigote forms) and were treated daily with PBS, meloxicam (0.5 mg/kg) or etoricoxib (0.6 mg/kg). The animals were sacrificed on the 21st day of infection and we collected the cardiac tissue and spleen cells for tissue culture. We observed that treatment with COX-2 inhibitors was able to decrease synthesis of PGE2 by spleen cells. This reduction was accompanied by reduction of the inflammatory infiltrate, parasite nets, cardiac fibrosis and fewer COX-2 positive cells in cardiac tissue obtained from these animals. In conclusion, treatment with COX-2 inhibitors, and consequent inhibition of PGE2 synthesis, was able to reduce the cardiac damage observed during the acute phase of experimental Chagas disease, thus demonstrating the involvement of this mediator in the cardiac lesion.

High-grade cervical intraepithelial neoplasia, human papillomavirus and factors connected with recurrence following surgical treatment.

Although effective strategies for preventing cancer of the uterine cervix exist, this disease continues to be a serious health problem worldwide, especially in developing countries. Today, the role of human papillomavirus (HPV) as a causal factor for the emergence of cervical cancer and its precursor lesions is well established, and prevention programs against cervical cancer are based on detecting cervical intraepithelial neoplasia (CIN). HPV present immunological evasion mechanisms that inhibit detection of the virus by the host, which may result in persistent chronic infection and irrevocably comprise the host defenses. Conization is the surgical technique most used for treating high-grade CIN, since it makes it possible to exclude invasive neoplasia, evaluate resection margins and preserve fertility. However, several factors have been considered to be indicators for residual disease. This review had the aim of covering some factors relating to persistence and recurrence of high-grade CIN following conization.

Inducible cyclooxygenase released prostaglandin mediates immunosuppression in acute phase of experimental Trypanosoma cruzi infection.

We investigated the possible role of prostaglandins produced by COX-2 in the immunosuppression observed during Trypanosoma cruzi infection. Con-A-stimulated splenocytes isolated from mice on days 5, 10, and 15 of infection released large amounts of PGE2 and this release was inhibited by the treatment of animals with sodium salicylate or meloxicam. The treatment of the animals with these drugs enhanced the release of IL-2 by splenocytes from T. cruzi-infected animals and significantly reduced the blood parasitemia and delayed the mortality of the infected mice. Furthermore, the release of TNF-alpha, IFN-gamma, IL-4, and IL-10 by Con-A-stimulated splenocytes obtained from infected mice on days 5, 10, and 15 of the infection was significantly inhibited by treatment of the animals with salicylate or meloxicam. In conclusion, the results suggest that the prostaglandins produced mainly by COX-2 mediate the immunosuppression observed in the acute phase of T. cruzi infection.

Pro-inflammatory effects of cholera toxin: role of tumor necrosis factor alpha.

Cholera toxin has been traditionally described as the one that does not induce inflammation. It has, however, potent adjuvant and immuno-modulatory activities. Since the adjuvanticity of other compounds is linked to their capacity to induce inflammation, in the present study the pro-inflammatory activity of cholera toxin was investigated. We studied this activity in the following rat models of inflammation: paw edema and neutrophil migration into the peritoneal cavity, and evaluated cholera toxin's effect on tumor necrosis factor alpha (TNF-alpha) production by mouse macrophages. We, also, explored the effects of dexamethasone (DEXA) and of two inhibitors of TNF-alpha production, thalidomide (TAL) and pentoxifylline, on paw swelling. Cholera toxin-induced significant and dose-dependent paw edema, which peaked 48 h after toxin challenge (Cholera toxin(2.5 microg): 2.39 +/- 0.22 ml). Cholera toxin B subunit did not show edematogenic activity. DEXA, TAL and pentoxifylline significantly reduced cholera toxin-induced edema (DEXA(0.5 mg/kg): 42.6% of inhibition; TAL(45 mg/kg): 36% of inhibition; pentoxifylline (45 mg/kg): 61% of inhibition). Neither cholera toxin nor its B subunit induced neutrophil migration into peritoneal cavities. Cholera toxin stimulated the release of TNF-alpha by macrophages (cholera toxin(10 microg): 11.46 +/- 0.44 UI/ml). These data provide evidences that cholera toxin exhibits significant pro-inflammatory activity. It also indicates the role of TNF-alpha upon the pathophysiology of this event based on the inhibitory action of DEXA, TAL and pentoxifylline, and on TNF-alpha secretion induced by cholera toxin.

Influence of the electric charge of the antigen and the immune complex (IC) lattice on the IC activation of human complement.

In order to understand the mechanism of complement (C) activation by immune complexes (ICs), the anti-complementary effect of ICs containing cationized antigens was compared in vitro to that using ICs formed by native antigens. ICs were prepared with affinity-purified rabbit polyclonal IgG antibovine serum albumin (BSA) antibody and either native BSA (isoelectric point 4.2) or BSA rendered cationic by treatment with ethylenediamine (isoelectric point 9.4). Native and cationized antigens were characterized by isoelectric focusing. ICs containing anti-BSA IgG or F(ab')2, formed either at equivalence or in excess of native or cationized antigen, were submitted to ultracentrifugation in a sucrose gradient for mesh size determination. The anti-complementary effect of ICs was evaluated by kinetic determination of haemolytic activity of human serum on haemolysin-sensitized sheep red blood cells. In conditions of antigen excess, the ICs formed by cationized BSA were significantly more efficient in activating human complement than those formed by native antigen. This higher activity was dependent on cationized antigen complexed with complete antibody molecules, as non-complexed cationized BSA or ICs prepared with F(ab')2 fragments were inactive under the same experimental conditions. Furthermore, this difference did not depend on the mesh size of the immune complexes. Our results suggest that the balance between antigen, antibody and C may be of importance in vivo for the onset and course of infections and other pathological processes involving IC formation. ICs containing cationized antigens should be proven of value in experimental models for studies on the regulation of C activation.

Involvement of prostaglandins in the immunosuppression occurring during experimental infection by Paracoccidioides brasiliensis.

We investigated whether PGE2 mediates the immunosuppression observed during Paracoccidioides brasilensis infection. Con-A-stimulated splenocytes, isolated from mice on days 15 and 60 of infection, release high amounts of PGE2, this release was inhibited by the treatment of animals with indomethacin, sodium salicylate or meloxicam. The treatment of the animals with salicylate or meloxicam, but not indomethacin, enhanced the release of IL-2 by splenocytes from animals on day 15, but not on day 60 of infection. Furthermore, we demonstrated that the productions of TNF-alpha, IFN-gamma, IL-4, and IL-10 by Con-A-stimulated splenocytes from mice at 15 days of infection were inhibited by treatment with salicylate or meloxicam. Indomethacin inhibited only TNF-alpha and IFN-gamma production. The three treatments caused reduction of granuloma areas in the liver and lungs of infected mice. In conclusion, results suggest that the PGE2 released by COX-2 mediates the immunosuppression early on (day 15), but not during the later phase (60 days) of P. brasiliensis infection by a mechanism dependent upon IL-4 and IL-10.

Role of lipocortin-1 in the anti-hyperalgesic actions of dexamethasone.

1. The effect of dexamethasone, lipocorton-1(2-26) and an antiserum to lipocortin-1(2-26) (LCPS1) upon the hyperalgesic activities in rats of carrageenin, bradykinin, tumour necrosis factor alpha (TNF alpha), interleukin-1(2), interleukin-6 (IL-6), interleukin-8 (IL-8), prostaglandin E beta (PGE2) and dopamine were investigated in a model of mechanical hyperalgesia. 2. Hyperalgesic responses to intraplantar (i.pl.) injections of carrageenin (100 micrograms), bradykinin (500 ng), TNF alpha (2.5 pg), IL-1 beta (0.5 pg), and IL-6 (1.0 ng), but not responses to IL-8 (0.1 ng), PGE2 (100 ng) and dopamine (10 micrograms), were inhibited by pretreatment with dexamethasone (0.5 mg kg-1, subcutaneously, s.c., or 0.04-5.0 micrograms/paw). 3. Inhibition of hyperalgesic responses to injections (i.pl.) of bradykinin (500 ng) and IL-1 beta (0.5 pg) by dexamethasone (0.5 mg kg-1, s.c.) was reversed by LCPS1 (0.5 ml kg-1, injected s.c., 24 h and 1 h before hyperalgesic substances) and hyperalgesic responses to injections (i.pl.) of bradykinin (500 ng), TNF alpha (2.5 pg) and IL-1 beta (0.5 pg), but not responses to PGE2 (100 ng), were inhibited by pretreatment with lipocortin-1(2-26) (100 micrograms/paw). Also, lipocortin-1(2-26) (30 and 100 micrograms ml-1 and dexamethasone (10 micrograms ml-1) inhibited TNF alpha release by cells of the J774 (murine macrophage-like) cell-line stimulated with LPS (3 micrograms ml-1), and LCPS1 partially reversed the inhibition by dexamethasone. These data are consistent with an important role for endogenous lipocortin-1(2-26) in mediating the anti-hyperalgesic effect of dexamethasone, with inhibiton of TNF alpha production by lipocortin-1(2-26) contributing, in part, to this role. 4. Although arachidonic acid by itself was not hyperalgesic, the hyperalgesic response to IL-1 beta (0.25 pg, i.pl.) was potentiated by arachidonic acid (50 micrograms) and the potentiated response was inhibited by dexamethasone (50 micrograms, i.pl.) and lipocortin-1(2-26) (100 micrograms, i.pl.). Also, lipocortin-1(2-26) (30 and 100 micrograms ml-1) inhibited/abolished PGE2 release by J774 cells stimulated with LPS (3 micrograms ml-1). These data suggest that, in inflammatory hyperalgesia, inhibition of the induction of cyclo-oxygenase 2 (COX-2), rather than phospholipase A2, by dexamethasone and lipocortin-1(2-26) accounts for the anti-hyperalgesic effects of these agents. 5. The above data support the notion that induction of lipocortin by dexamethasone plays a major role in the inhibition by dexamethasone of inflammatory hyperalgesia evoked by carrageenin, bradykinin and the cytokines TNF alpha, IL-1 beta and IL-6, and provides additional evidence that the biological activity of lipocortin resides within the peptide lipocortin-1(2-26). Further, the data suggest that inhibition of lipocortin-1(2-26) of eicosanoid production by COX-2 also contributes to the anti-hyperalgesic effect of lipocortin-1.