Local Immunomodulation Research Articles

Role of gastroesophageal reflux disease in lung transplantation.

Lung transplantation is one of the highest risk solid organ transplant modalities. Recent studies have demonstrated a relationship between gastroesophageal reflux disease (GERD) and lung transplant outcomes, including acute and chronic rejection. The aim of this review is to discuss the pathophysiology, evaluation, and management of GERD in lung transplantation, as informed by the most recent publications in the field. The pathophysiology of reflux-induced lung injury includes the effects of aspiration and local immunomodulation in the development of pulmonary decline and histologic rejection, as reflective of allograft injury. Modalities of reflux and esophageal assessment, including ambulatory pH testing, impedance, and esophageal manometry, are discussed, as well as timing of these evaluations relative to transplantation. Finally, antireflux treatments are reviewed, including medical acid suppression and surgical fundoplication, as well as the safety, efficacy, and timing of such treatments relative to transplantation. Our review of the data supports an association between GERD and allograft injury, encouraging a strategy of early diagnosis and aggressive reflux management in lung transplant recipients to improve transplant outcomes. Further studies are needed to explore additional objective measures of reflux and aspiration, better compare medical and surgical antireflux treatment options, extend follow-up times to capture longer-term clinical outcomes, and investigate newer interventions including minimally invasive surgery and advanced endoscopic techniques.

Biopatterned CTLA4/Fc Matrices Facilitate Local Immunomodulation, Engraftment, and Glucose Homeostasis After Pancreatic Islet Transplantation.

Pancreatic islet transplantation (PIT) represents a potential therapy to circumvent the need for exogenous insulin in type 1 diabetes. However, PIT remains limited by lack of donor islets and the need for long-term multidrug immunosuppression to prevent alloimmune islet rejection. Our goal was to evaluate a local immunoregulatory strategy that sustains islet allograft survival and restores glucose homeostasis in the absence of systemic immunosuppression. Nanogram quantities of murine CTLA4/Fc fusion protein were controllably delivered within human acellular dermal matrix scaffolds using an inkjet-based biopatterning technology and cotransplanted with allogeneic islets under the renal capsule to create an immunoregulatory microenvironment around the islet allograft. We achieved long-term engraftment of small loads of allogeneic islet cells with 40% of MHC-mismatched mouse recipients maintaining sustained normoglycemia following pancreatic β-cell ablation by streptozotocin. Biopatterned CTLA4/Fc local therapy was associated with expansion of Foxp3+ regulatory T cells and shifts in cytokine production and gene expression from proinflammatory to regulatory profiles, thus substantially benefiting islet allografts survival and function. This study is a new paradigm for targeted therapies in PIT that demonstrates the favorable effects of immune alterations in the transplant milieu and suggests a unique strategy for minimizing systemic immunosuppression and promoting islet allograft survival.

Glucocorticoid receptor haplotypes conferring increased sensitivity (BclI and N363S) are associated with faster progression of multiple sclerosis

As high cortisol levels are implicated in suppressed disease activity of multiple sclerosis (MS), glucocorticoid receptor (GR) polymorphisms that affect glucocorticoid (GC) sensitivity may impact on this by changing local immunomodulation or regulation of the hypothalamus–pituitary–adrenal (HPA)-axis. In this post-mortem study, we investigated whether GR haplotypes affect MS disease course and production of cortisol and soluble CD163 (sCD163), a molecule induced by GC on microglia/macrophages. We found that GR haplotypes that confer high GC sensitivity are associated with more aggressive MS but do not affect levels of cortisol secreted by the HPA-axis or shedding of CD163.

Влияние локального применения рекомбинантных цитокинов на течение и исход дентальной имплантации с закрытым синус-лифтингом

<p class="p1"><span class="s1">In order to increase the bone height in the distal portions of the maxilla during dental implantation the method of «closed» sinus lift is widely applied. Manipulation is considered to be a minimally invasive osteotomy but despite this fact, it is associated with a high complication rate that might lead to the loss of the dental implant. </span></p><p class="p1"><span class="s1">The purpose of the project is to analyze the evaluation of the effectiveness of immunomodulators local application during the "closed" sinus lift manipulation that is based on local in uence of «Betaleukin» and «Roncoleukin» drugs in the area of unexpected perforation of the mucous membrane of the maxillary sinus during the formation of the bone bed and close contact surface of the implant with a damaged part. A clinical randomized trial was held which of 105 patients aged 30 to 57, divided into three groups: comparison group, n = 35 (implants placed by the closed sinus lift standard method); betaleukin group, n = 35 (in addition to the surgery performed local recombinant IL-1β immunotherapy was conducted); roncoleukin group, n = 35 (the surgery done was combined with the local ronkoleukine immunomodulation). Within 10-12 days after the surgery hydrogel «Koleteks-Beta» and a gel containing recombinant IL-2 for a local effect on the oral mucous membrane around the installed implant were applied. During the study, all patients had a number of clinical signs that allow assessing the dynamics of post-operative course of the integration process of implants. Radiological control in the postoperative period was conducted to evaluate the position of the implant in relation to the maxillary sinus; the dynamics of bone tissue around it was observed. It was found out that the local treatment by immunomodulators improves the performance dynamics of clinical postoperative period; it reduces the risk of graft rejection and increases the ef ciency of integration process. </span></p>

Non-hepatic Islet transplantation: Local Delivery of Reagents to Aid in Islet Engraftment

Abstract Islet transplantation offers a potential cell replacement treatment for type 1 diabetes, but faces a number of significant challenges, including: 1) poor short-term islet survival and 2) long term islet loss by alloimmunity and recurring autoimmunity. Method We have developed a Bioengineered Islet Implant (BII) device that incorporates local biological reagent delivery to promote islet immune tolerance and survival. The BII consists of two integrated components: a poly-vinyl-alcohol (PVA) scaffold to support islets within a collagen hydrogel, and a local drug delivery mechanism located within the implant device through incorporating drug-alginate delivery system. Results The implant device is capable of reversing STZ-induced diabetes when implanted in mesentery or subcutaneous sites. Short-term islet survival measured at 24 hours is enhanced in recipient transplants utilizing BII incorporating local delivery of VEGF. As proof-of-concept for delivery of local immunomodulators together with transplanted islets, we incorporated anti-CD3e MAb (2C11) or anti-CD95 MAb (Jo-2) within the BII, and respectively found dramatically reduced lymphocyte infiltration into islets and a delay in the onset of diabetes in the DO11/RIP-Ova murine model of T1D. In addition, local delivery within the BII of the extracellular component hyaluronic acid led to the generation of enhanced antigen-specific IL-10 responses. Conclusion These data demonstrate the potential for a novel strategy of islet cytoprotection during transplantation, providing a suitable matrix and local immunomodulation capable of supporting islet cell engraftment, survival, and function in non-hepatic sites without encapsulation.

Pancreatic cancer microenvironment, to target or not to target?

We have collectively been spoiled by the astounding clinical benefit of antimicrobials. Much like the discovery and use of penicillin to eradicate once deadly infections, we continue to desperately search for the next “magic bullet” to kill cancer while sparing the non‐transformed cells. Greater appreciation for the molecular intricacies of malignancy has resulted in dedicated pursuit of cancer genomics and large‐scale informatics to identify “drugable” targets within the cancer cell itself. However, studies at the bench elucidating a dynamic relationship between tumor and microenvironment have become more common and demonstrate promise for novel therapeutic intervention.

Emerging Implications for Extracellular Matrix-Based Technologies in Vascularized Composite Allotransplantation.

Despite recent progress in vascularized composite allotransplantation (VCA), limitations including complex, high dose immunosuppression regimens, lifelong risk of toxicity from immunosuppressants, acute and most critically chronic graft rejection, and suboptimal nerve regeneration remain particularly challenging obstacles restricting clinical progress. When properly configured, customized, and implemented, biomaterials derived from the extracellular matrix (ECM) retain bioactive molecules and immunomodulatory properties that can promote stem cell migration, proliferation and differentiation, and constructive functional tissue remodeling. The present paper reviews the emerging implications of ECM-based technologies in VCA, including local immunomodulation, tissue repair, nerve regeneration, minimally invasive graft targeted drug delivery, stem cell transplantation, and other donor graft manipulation.

Surgical meshes coated with mesenchymal stem cells provide an anti-inflammatory environment by a M2 macrophage polarization

Surgical meshes are widely used in clinics to reinforce soft tissue’s defects, and to give support to prolapsed organs. However, the implantation of surgical meshes is commonly related with an inflammatory response being difficult to eradicate without removing the mesh. Here we hypothesize that the combined use of surgical meshes and mesenchymal stem cells (MSCs) could be a useful tool to reduce the inflammatory reaction secondary to mesh implantation.In vitro determinations of viability, metabolic activity and immunomodulation assays were performed on MSCs-coated meshes. Magnetic resonance imaging, evaluation by laparoscopic optical system and histology were performed for safety assessment. Finally, flow cytometry and qRT-PCR were used to elucidate the mechanism of action of MSCs-coated meshes.Our results demonstrate the feasibility to obtain MSCs-coated surgical meshes and their cryopreservability to be used as an ‘off the shelf’ product. These biological meshes fulfill the safety aspects as non-adverse effects were observed when compared to controls. Moreover, both in vitro and in vivo studies demonstrated that, local immunomodulation of implanted meshes is mediated by a macrophage polarization towards an anti-inflammatory phenotype.In conclusion, the combined usage of surgical meshes with MSCs fulfills the safety requirements for a future clinical application, providing an anti-inflammatory environment that could reduce the inflammatory processes commonly observed after surgical mesh implantation. Statement of SignificanceSurgical meshes are medical devices widely used in clinics to resolve hernias and organs’ prolapses, among other disorders. However, the implantation of surgical meshes is commonly related with an inflammatory response being difficult to eradicate without removing the mesh, causing pain and discomfort in the patient. Previously, the anti-inflammatory, immunomodulatory and pro-regenerative ability of mesenchymal stem cells (MSCs) have been described.To our knowledge, this is the first report where the anti-inflammatory and pro-regenerative ability of MSCs have been successfully applied in combination with surgical meshes, reducing the inflammatory processes commonly observed after mesh implantation. Moreover, our in vitro and in vivo results highlight the safety and efficacy of these bioactive meshes as a ‘ready to use’ medical product.

Giant extragenital Bowen’s disease

Giant extragenital forms of Morbus Bowen are extremely rare. The already described cases in the word literature are most commonly with periungual localization, as well as located on the foot and neck area. The clinical manifestation is presented most commonly by non-specific erythematous to erythematous-squamous plaques or papules, which is confusing to the clinician. From the pathogenic point of view, it is important to be confirmed or rejected the presence of human papilloma viruses (HPVs) in each case of affected patient, as this information is mandatory in respect to the adequate selection of the subsequent regimen. If HPVs are detected, systemic antiviral therapy could be initiated to reduce the size of the lesions before subsequent surgical eradication. A postoperative prevention through vaccination could be also considered additionally. In cases of HPV-negative giant extragenital forms of Morbus Bowen (as in the described patient), the focus should be on local immunomodulation by substances such as imiquimod, which reduce the size of the lesions, thereby creating optimal opportunities for their future surgical eradication. Other possible options described in the literature include topical application of 5-fluorouracil, photodynamic therapy, cryotherapy, and laser therapy (carbon dioxide laser). The choice of the most appropriate regimen should have been an individual decision of the clinician, considering also the location and the extent of the lesion.

Interleukin-12 Immunomodulation Delays the Onset of Lethal Peritoneal Disease of Ovarian Cancer.

The omental fat band (OFB) is the predominant site for metastatic seeding of ovarian cancer. Previously, we highlighted the influx and accumulation of neutrophils and macrophages in the OFB following syngeneic ovarian cancer cell seeding as an important factor in the development of a protumorigenic cascade. Here we investigated localized immunomodulation as a means of promoting a successful protective response. As an important TH1-type immunomodulator, interleukin (IL)-12 has previously been investigated clinically as an anticancer therapeutic. However, systemic IL-12 administration was associated with serious side effects, galvanizing the development of immune or accessory cells engineered to express secreted or membrane-bound IL-12 (mbIL-12). Using an mbIL-12-expressing cell variant, we demonstrate that localized IL-12 in the tumor microenvironment significantly delays disease development. The mbIL-12-mediated decrease in tumor burden was associated with a significant reduction in neutrophil and macrophage infiltration in the OFB, and correlated with a reduced expression of neutrophil and macrophage chemoattractants (CXCL1, -2, -3 and CCL2, -7). Vaccination with mitotically impaired tumor cells did not confer protection against subsequent tumor challenge, indicating that IL-12 did not impact the immunogenicity of the cancer cells. Our findings are in agreement with previous reports suggesting that IL-12 may hold promise when delivered in a targeted and sustained manner to the omental microenvironment. Furthermore, resident cells within the omental microenvironment may provide a reservoir that can be activated and mobilized to prevent metastatic seeding within the peritoneum and, therefore, may be targets for chemotherapeutics.

Feasibility of localized immunosuppression: 1. Exploratory studies with glucocorticoids in a biohybrid device designed for cell transplantation.

Emerging biotechnologies, such as the use of biohybrid devices for cellular therapies, are showing increasing therapeutic promise for the treatment of various diseases, including type 1 diabetes mellitus. The functionality of such devices could be greatly enhanced if successful localized immunosuppression regimens could be established, since they would eliminate the many otherwise unavoidable side effects of currently used systemic immunosuppressive therapies. The existence of local immune privilege at some specialized tissues, such as the eye, CNS, or pregnant uterus, supports the feasibility of localized immunomodulation, and such an approach is particularly well-suited for cell transplant therapies where all transplanted tissue is localized within a device. Following the success of syngeneic transplantation in a subcutaneous prevascularized device as a bioartificial pancreas in a rodent model, we now report the first results of exploratory in vivo islet allograft studies in rats using locally delivered glucocorticoids (dexamethasone phosphate and the soft steroid loteprednol etabonate). Following in vitro assessments, in silico drug distribution models were used to establish tentative therapeutic dose ranges. Sustained local delivery was achieved via implantable osmotic mini-pumps through a central sprinkler, as well as with a sustained-delivery formulation for loteprednol etabonate using poly(D,L-lactic) acid (PLA) microspheres. Doses delivered locally were approximately hundred-fold smaller than those typically used in systemic treatments. While several solubility, stability, and implantation problems still remain to be addressed, both compounds showed promise in their ability to prolong graft survival after tapering of systemic immunosuppression, compared to control groups.

Иммунологические аспекты локальной иммуномодуляции мукозальной вакциной системного действия у больных хроническим тонзиллитом и после тонзиллэктомии

Иммунологические аспекты локальной иммуномодуляции мукозальной вакциной системного действия у больных хроническим тонзиллитом и после тонзиллэктомии

ULCERAÇÕES GENITAIS HERPÉTICAS RECALCITRANTES TRATADAS COM IMIQUIMOD ASSOCIADO AO VALACICLOVIR

O vírus herpes simplex é o principal responsável por ulcerações genitais de etiologia infecciosa. No contexto de imunodeficiência, manifesta-se frequentemente de forma atípica, com lesões mais floridas, sendo também frequente a resistência à terapêutica clássica, resultando numa evolução prolongada.Descreve-se o caso clinico de um homem de 25 anos, com antecedentes de imunodeficiência comum variável, que recorre à nossa consulta por ulcerações genitais extensas e exuberantes da região inguino-escrotal de etiologia herpética. Após terapêutica prolongada com valaciclovir, o doente mantinha-se sintomático, tendo-se optado pela associação de imiquimod 5% tópico, com resolução do quadro clínico e excelente tolerância.O imiquimod, funcionando como um imunomodulador local, estimula a resposta imunológica antiviral. Configura- -se assim como uma alternativa nos casos de infeções herpéticas de difícil controlo, especialmente em doentes imunodeprimidos.

Effect of uterine immunomodulation on hematobiochemical parameters in cyclic non-breeding cows

Aim: To study the effect of uterine immunomodulation on hematobiochemical parameters and total immunoglobulin concentration in cyclic non-breeding cows. Materials and Methods: Twenty-one repeat breeding cows around Bhubaneswar area were screened by white side test to detect and treat the endometritis and were assigned to three treatment protocols with an equal number of seven animals in each group. Cows in control group were administered with 50 ml of normal saline while treatment Group I animals were given single intrauterine infusion of 20 ml of fresh colostrum and treatment Group II animals received nonpathogenic Escherichia coli in 10 ml sterile saline. The blood samples were collected from all the experimental animals, and hematobiochemical parameters and total immunoglobulin concentration were estimated. Results: A high significant difference (p<0.01) was accounted in lymphocyte count of E. coli treated group within different days of sampling. Analysis of variance recorded a highly significant difference with neutrophil percent in E. coli lavaged cows. In colostrum treated group monocyte count showed a significant difference (p<0.01) between 0 and 14 th day of sampling. The analysis of hematocrit values did not show any significant difference apart from the erythrocyte sedimentation rate parameter in the colostrum infused group with the highest significant (p<0.01) variation being observed between 7 th and 14 th day of sampling. The analysis of aspartate amino transferase values in the colostrum lavaged cows revealed a significant difference, but that of alanine amino transferase values did not show any significant difference. Comparison of immunoglobulin values for different days in all the treatment protocol revealed a highly significant (p<0.05) difference within various days of sampling. Conclusion: In the present study, the local immunomodulation by different agents have been highlighted and which indicated potentiation of uterine immunity by different drug that might serve as a new direction of treatment to uterine diseases. The scope of research in the future should be widened by considering a larger population for validation.

Mesenchymal Stem Cell Therapy Promotes Corneal Allograft Survival in Rats by Local and Systemic Immunomodulation

Mesenchymal stem cells (MSCs) are being investigated extensively due to their ability to dampen immune responses. Here, we tested the ability of MSCs from three distinct sources to prolong rat corneal allograft survival. A fully allogeneic rat cornea transplant model (DA to LEW) was used. Recipient rats received 1 × 10(6) MSCs (syn [LEW], allo [DA] or third-party [Wistar Furth]) intravenously 7 days before transplantation and again on the day of transplantation (day 0). A high percentage of untreated and syn-MSC treated allografts were rejected (80% and 100%, respectively). Preactivation of syn-MSCs with interferon gamma also failed to prolong allograft survival. Conversely, corneal allograft survival was significantly prolonged in allo-MSC treated (90%) and third-party MSC treated (80%) allograft recipients. Flow cytometric analysis revealed less infiltrating natural killer T cells in corneas of both allo- and third-party MSC treated animals, coupled with a higher proportion of splenic CD4+Foxp3+ regulatory T cells, compared to controls. In the case of allo- and third-party MSCs, results from a delayed-type hypersensitivity assay clearly showed that hypo-responsiveness was specific for corneal donor-associated allo-antigens. Thus, allo- and third-party MSC treatment prolongs corneal allograft survival by suppressing peripheral immune responses and promoting an intragraft immunoregulatory milieu.

IMA901 for metastatic renal cell carcinoma in the context of new approaches to immunotherapy.

The promising option of immunotherapy for metastatic renal cell carcinoma has evolved from rather unspecific approaches to a specific activation of an anti-tumor T-cell response. The latest step is a synthetic peptide vaccine called IMA901, which demonstrated a clear association between a provoked T-cell response and a prolonged overall survival. The results of IMA901 for the treatment of metastatic renal cell carcinoma are discussed together with new approaches to immunotherapy, such as local and systemic immunomodulation with adjuvants, checkpoint inhibitors, classical chemotherapeutics, such as cyclophosphamide or tyrosine kinase inhibitors. The capability of theses substances to modulate leukocytes subsets, such as myeloid-derived suppressor cells, Tregs or Th17 cells, are outlined together with the possibility to combine them with tumor vaccination strategies to achieve a higher cancer specificity and immunogenicity.

HPV16 E2 protein promotes innate immunity by modulating immunosuppressive status

The balance between active immune responses against human papillomavirus (HPV) and HPV-induced immune escape regulates viral clearance and carcinogenesis. To understand the role of the early viral protein HPV16 E2 in host innate immune responses, the HPV16 E2-transfected murine squamous cell carcinoma cell line SCCVII (SCC/E2) was generated and anti-tumor responses in T-cell-depleted mice were evaluated. Tumor growth of SCC/E2 was markedly reduced. Cytotoxicity against the NK-sensitive targets YAC-1 and SCCVII was clearly enhanced in SCC/E2-inoculated mice. Despite the comparable ratio of NK cells, the proportion of CD11b+Gr-1+ myeloid-derived suppressor cells (MDSCs) was significantly decreased in SCC/E2-inoculated mice. The transcription of MDSC-related mediators such as inducible nitric oxide synthase, indoleamine 2,3-dioxygenase, and heme oxygenase-1 was significantly impaired in the SCC/E2-inoculated tumor tissues on day 3. Our results suggest that HPV16 E2 promotes anti-tumor innate effector function by modulating immunoregulatory events mediated by MDSCs and their mediators. This report describes a new role for HPV16 E2 as a local immunomodulator at infected sites.

Depleting tumor-specific Tregs at a single site eradicates disseminated tumors

Activation of TLR9 by direct injection of unmethylated CpG nucleotides into a tumor can induce a therapeutic immune response; however, Tregs eventually inhibit the antitumor immune response and thereby limit the power of cancer immunotherapies. In tumor-bearing mice, we found that Tregs within the tumor preferentially express the cell surface markers CTLA-4 and OX40. We show that intratumoral coinjection of anti-CTLA-4 and anti-OX40 together with CpG depleted tumor-infiltrating Tregs. This in situ immunomodulation, which was performed with low doses of antibodies in a single tumor, generated a systemic antitumor immune response that eradicated disseminated disease in mice. Further, this treatment modality was effective against established CNS lymphoma with leptomeningeal metastases, sites that are usually considered to be tumor cell sanctuaries in the context of conventional systemic therapy. These results demonstrate that antitumor immune effectors elicited by local immunomodulation can eradicate tumor cells at distant sites. We propose that, rather than using mAbs to target cancer cells systemically, mAbs could be used to target the tumor infiltrative immune cells locally, thereby eliciting a systemic immune response.

Depleting tumor-specific Tregs at a single site eradicates disseminated tumors

Activation of TLR9 by direct injection of unmethylated CpG nucleotides into a tumor can induce a therapeutic immune response; however, Tregs eventually inhibit the antitumor immune response and thereby limit the power of cancer immunotherapies. In tumor-bearing mice, we found that Tregs within the tumor preferentially express the cell surface markers CTLA-4 and OX40. We show that intratumoral coinjection of anti-CTLA-4 and anti-OX40 together with CpG depleted tumor-infiltrating Tregs. This in situ immunomodulation, which was performed with low doses of antibodies in a single tumor, generated a systemic antitumor immune response that eradicated disseminated disease in mice. Further, this treatment modality was effective against established CNS lymphoma with leptomeningeal metastases, sites that are usually considered to be tumor cell sanctuaries in the context of conventional systemic therapy. These results demonstrate that antitumor immune effectors elicited by local immunomodulation can eradicate tumor cells at distant sites. We propose that, rather than using mAbs to target cancer cells systemically, mAbs could be used to target the tumor infiltrative immune cells locally, thereby eliciting a systemic immune response.

Killer Treg cells ameliorate inflammatory insulitis in non-obese diabetic mice through local and systemic immunomodulation

Treg cells endowed with enhanced killing activity through decoration with Fas-ligand (FasL) protein (killer Treg) have been effective in delay of hyperglycemia in prediabetic non-obese diabetic (NOD) mice. In this study, we assessed the therapeutic efficacy of these cells, harvested from age-matched euglycemic NOD donors, on the course of disease in new-onset diabetics. One dose of 4 × 10(6) killer Treg cells stabilized blood glucose associated with increased insulin levels in 5 of 9 mice and partially reversed the severity of islet inflammation, whereas naive Treg cells did not modulate the course of disease significantly. Killer Treg cells were shown to operate through induction of cell apoptosis within the pancreatic lymph nodes, resulting in reduced efficiency of adoptive disease transfer to NOD/SCID recipients. A second mechanism of action consisted of increased fractions of CD4(+)CD25(-)FoxP3(+) T cells in the pancreas and all lymphoid organs. Immunomodulation with FasL rather than Treg cells enhanced the expression of CD25 and FoxP3 in the thymus, suggesting a possible contribution of thymic output to prolonged stabilization of the glucose levels. Autologous Treg cells evolve as excellent vehicles for targeted delivery of FasL as an immunomodulatory protein, which delete pathogenic cells at the site of inflammation and induce systemic dominance of suppressor subsets.