Anti-tumoral Effects of Recombinant Lactococcus lactis Strain Secreting IL-17A Cytokine.

Anti-tumoral Effects of Recombinant Lactococcus lactis Strain Secreting IL-17A Cytokine.

Interleukin-17A (IL-17A) is a pro-inflammatory cytokine produced by T H 17 cells that participate in and contribute to host defense and autoimmune diseases. We recently reported the antitumor properties of the probiotic strain Lactobacillus casei BL23 and TH17 cells in mice was shown to play an important role in this beneficial effect.

In order to better understand the role of IL-17A in cancer, we construct strain recombinant of Lactococcus lactis produce this cytokine and we are determined biological activity in: (i) the test bioassay for the induction of IL-6 production of fibroblasts murine 3T3 L1 cell line and (ii) The allograft mouse models of human papilloma virus (HPV) induced cancer.

Our data indicate that the recombinant L. lactis produce and efficiently secrete biologically active IL-17A cytokine. Interestingly, ~ 26% of mice treated with L. lactis intranasal-IL-17A and challenged with TC-1 cells remained tumor free through an experiment, in contrast to the control mice treated with wild strains of L. lactis developed 100% tumor is aggressive.

In addition, the median size of ~ 74% tumor-bearing mice treated with recombinant L. lactis-IL-17A, significantly lower than mice treated with L. lactis-wt. Overall, our results indicate that the intranasal administration of L. lactis secreting IL-17A results in partial protection against TC-1 tumor-induced in rats, confirming the antitumor effects of this cytokine is in our cancer model.

Anti-tumoral Effects of Recombinant Lactococcus lactis Strain Secreting IL-17A Cytokine.
Anti-tumoral Effects of Recombinant Lactococcus lactis Strain Secreting IL-17A Cytokine.

Anti-colorectal cancer effects of anti-p21Ras scFv delivered by the recombinant adenovirus KGHV500 and cytokine-induced killer cells.

BACKGROUND
Colorectal cancer (CRC) is the most common type of gastrointestinal cancer. CRC adenovirus-mediated gene therapy holds great promise for the treatment of malignancies. However, the intravenous delivery of adenovirus limited exhibit anti-tumor activity in vivo when used alone.


METHOD
In this study, the antitumor activity of recombinant adenovirus KGHV500 assessed by MTT, TUNEL, Matrigel invasion and cell migration test. To improve the intravenous delivery KGHV500 vivo cytokine-induced killer (CIK) cells are used as vectors to carry KGHV500 second. We explored whether CIK cells can carry KGHV500 recombinant adenovirus containing anti-p21Ras single chain variable antibody fragment (scFv) gene into the tumor and increase the antitumor potential.


RESULTS
Our results indicate that KGHV500 showed significant antitumor activity in vitro. In mouse xenograft tumor model SW480 nude, combined with KGHV500 CIK cells can induce a higher antitumor activity against colorectal cancer in vivo than that caused by either CIK or KGHV500 only. After seven days of treatment, adenovirus and scFv was detected in tumor tissue but not in normal tissues was detected by immunohistochemistry. Therefore, KGHV500 replicates in tumor and anti-p21Ras managed to express scFv in a xenograft model of colorectal cancer.


CONCLUSION
Our study provides a new strategy for the treatment of colorectal cancer by combining the CIK cells with recombinant adenovirus KGHV500 conducted scFv anti-p21 Ras.

Recombinant horse interleukin-4 and interleukin-10 induced a mixed inflammatory cytokine response in horse peripheral blood mononuclear cells.

Recombinant horse interleukin-4 and interleukin-10 induced a mixed inflammatory cytokine response in horse peripheral blood mononuclear cells.

Background and Objectives
Most Interleukin (IL) -4 and IL-10 activates immune cells and induce a humoral immune response. However, horses recombinant version of IL-4 and IL-10 has not been studied to understand their immunomodulating activities.

This study aims to produce horses mature recombinant IL-4 and IL-10 in Escherichia coli. Immune-modulating activity of recombinant equine IL-4 and IL-10 were investigated in peripheral blood mononuclear cells (PBMC).


Equine PBMC stimulated with recombinant IL-4 and IL-10. A PBMC proliferation was measured by XTT assay and induction of cytokines were measured by enzyme-linked immunosorbent assay and real-time polymerase chain reaction.

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of a molecular weight of 15 kDa shown for IL-4 and 19 kDa for IL-10. Recombinant IL-4 and IL-10 significantly induced cell proliferation at 250 ng / ml.

Results showed that IL-4 increased the expression of interferon-gamma (IFN-γ), IL-6, tumor necrosis factor-alpha (TNF-α), and IL-10, while the horse of recombinant IL-10 induced expression of IL-6, IFN-γ and TNF-α.
Conclusion
This study indicates that equine biologically active IL-4 and IL-10 can be produced in E. coli.

Recombinant horse interleukin-4 and interleukin-10 induced a mixed inflammatory cytokine response in horse peripheral blood mononuclear cells.
Recombinant horse interleukin-4 and interleukin-10 induced a mixed inflammatory cytokine response in horse peripheral blood mononuclear cells.

SOCS4 expressed by recombinant HSV protects against cytokine storm in a mouse model.

Oncolytic viruses are genetically engineered viruses are designed for the treatment of solid tumors, and is often combined with antitumor immunity of the host.

Challenges using oncolytic herpes simplex virus (oHSV) as a potent oncolytic agent is a potential host tissue damage caused by the production of various cytokines following oHSV intratumoral injection. HSV-suppressor of cytokine signaling 4 (SOCS4) recombinant virus was created to investigate whether it inhibits cytokine storm.

Recombinant HSV-SOCS4 and HSV-1 (F) is used to infect mice, and the level of representation of several cytokines, including monocyte chemoattractant protein-1, interleukin (IL) -1β, tumor necrosis factor-α, IL-6 and interferon-γ, in serum and bronchoalveolar lavage fluid (Balf) of infected mice is determined, and the immune cells in the spleen Balf and enumerated. lung damage, viral titers in the lungs, body weight and survival rate of infected mice was also determined and compared between the two groups.

The concentrations of cytokines rat-infected with HSV-SOCS4 significantly decreased compared to HSV-1 (F) -infected mice in Balf and serum, and a small cluster of differentiation (CD) Balf 11b + cells, and CD8 + CD62L + T cells and CD4 cells + CD62L + T spleen also been identified in mice with HSV-SOCS4-infected.

SOCS4 HSV-infected mice exhibited mild lung damage, a decrease in body weight loss and survival rate of 100%. The results of this study show that the regulator may SOCS4 protein useful for inhibiting cytokine overproduction, and that HSV-SOCS4 may provide a possible solution to control cytokine storm and its consequences after induction with oncolytic virus treatment.

In vitro spleen cell cytokine responses of adult mice immunized with a recombinant PorA (major outer membrane protein [MOMP]) from Campylobacter jejuni.

In vitro spleen cell cytokine responses of adult mice immunized with a recombinant PorA (major outer membrane protein [MOMP]) from Campylobacter jejuni.

No information on the cytokine profile to be used as a marker of Campylobacter jejuni infection protection. For this study, we used the outer membrane protein (MOMP [Pora]) as a vaccine for the protection and spleen cell cytokine as a marker of protection.

We cloned and expressed Pora from C. jejuni111 and mice immunized with intraperitoneal route. Subsequently, the mice orally challenged with C. jejuni 111.

live vaccine-induced protection as evidenced by a decrease in fecal excretion C. jejuni111. Cytokines were measured in vitro after stimulation of spleen cells with MOMP. Levels of pro-inflammatory cytokines, IL-12, TNF-α, IL-17A and IL-17F are similar in the control and test mice. Levels of pro-inflammatory cytokines, IL-2 and IFN-γ were higher in mice than in control mice test, and the level of pro-inflammatory cytokines, IL-8 and IL-1β was higher in the test mice than in control mice.

In between the two anti-inflammatory cytokines, the same level of IL-10 but higher for IL-4 in the test mice than in control mice. The ratio of pro-inflammatory anti-inflammatory cytokines showed a bias towards anti-inflammatory response in support of the production of antibodies that reflect the role of antibodies in immunity.

Cytokine patterns of production by spleen cells can be used as a marker of protection in a mouse model.

In vitro spleen cell cytokine responses of adult mice immunized with a recombinant PorA (major outer membrane protein [MOMP]) from Campylobacter jejuni.
In vitro spleen cell cytokine responses of adult mice immunized with a recombinant PorA (major outer membrane protein [MOMP]) from Campylobacter jejuni.

A New Efficient Method for Production of Recombinant Antitumor Cytokine TRAIL and Its Receptor-Selective Variant DR5-B.

The cytokine TRAIL induces apoptosis in tumor cells of various origin without affecting normal cells. clinical trials of TRAIL receptors (DR4 and DR5) agonist (recombinant TRAIL or death receptor antibodies) have largely failed because most human tumors are resistant to them. Today, the second generation of targeted agents in TRAIL-R with improved efficiency have been developed. To this end, we have developed DR5-B, selective TRAIL variants interact with DR5. We have developed a new efficient method for the production of TRAIL and DR5-B uses this protein expression in Escherichia coli strain B. Shuffle protein fractions isolated from the cytoplasm of cells and purified to a high degree of homogeneity using metal-affinity and ion exchange chromatography. The results of the protein is 211 and 173 mg from one liter of cell culture for DR5-B and TRAIL, respectively, which significantly exceed the results obtained by other methods. DR5-B kill tumor cells of different origins is more efficient and faster than TRAIL. The resulting preparation can be used for the study of TRAIL signaling pathways and in preclinical and clinical trials as an antitumor agent.

How a cytokine in your brain regulates your body weight?

According to figures, there are more than 1.9 billion overweight people, about one-third of whom are obese. Obesity hides the possible dangers of multiple diseases, so successful anti-obesity treatments are desperately needed.

It is well-known that interleukin-6 (IL-6) is a pro-inflammatory molecule that’s part of the innate immune system. However, researchers in the University of Gothenburg in Sweden have discovered a link between IL-6 from the brain and obesity. Through experiments in mice and rats, they confirmed that IL-6 does influence the risk of obesity and where this effect happens in the mind.

Interestingly, the brain’s control and utilization of IL-6 could be different from different parts of the body. The investigators wanted to know what happens to the level of IL-6 in the brain after a diet that leads to obesity.

Throughout the experiment, the researchers fed rats and mice with high-calorie foods like fat and hot mix. All these rodents, like humans, opt to eat too much when they eat foods that are high-calorie.

Obese mouse with low IL-6 brain levels

They discovered that IL-6 was reduced in fat rats and mice, but only in a region of the brain called the parabrachial nucleus (lPBN).

To research whether the reduction in IL-6 is bad or good for the metabolic health of rodents, the group has very selectively reduced the degree of IL-6 in lPBN by viral inheritance. The result is that even for rodents with a healthy diet, their weight and fat increase.

Consequently, the researchers believed that a drop in lPBN IL6 levels in obesity may lead to metabolic dysfunction and weight reduction. Since body weight is the result of energy intake and energy expenditure, any dysfunctionality in either of the 2 branches of energy equilibrium leads to weight gain.

After additional study, the team discovered that IL-6 made by the parabrachial nucleus affects both branches, namely, it reduces food intake and increases energy expenditure, while the latter increases brown fat action, so the body’s energy is used to create heat or burn fat. Therefore, lowering the IL-6 amount in lPBN breaks the whole energy balance.

Early studies in the University of Gothenburg have demonstrated that serum IL-6 levels in obese and overweight men are elevated but mind IL-6 levels measured in cerebrospinal fluid have been reduced.

The researchers also found that the decrease in IL-6 connected with obesity has been just found in men. Female rats and mice have normal IL-6 levels. The study team is currently investigating why women are immune to obesity-related IL-6-driven dysfunction.

Recombinant human GM-CSF

Glycosilation is better in Pychia recmbinant GM-CSF and it will give less problems with Oedemia in treated patients.

For Dendritic cell research more oftern E. coli recombinant GM-CSF and IL-4 is used.

Use of Growth Factors and Other Cytokines for Treatment of Injuries During a Radiation Public Health Emergency.

Journal: Radiation research


Due to the threat of a radiological or nuclear incident that could impact citizens, the U.S. Department of Health and Human Services tasked the National Institute of Allergy and Infectious Diseases (NIAID) with identifying and funding early- to mid-stage medical countermeasure (MCM) development to treat radiation-induced injuries. Given that the body’s natural response to radiation exposure includes production of growth factors and cytokines, and that the only drugs approved by the U.S. Food and Drug Administration to treat acute radiation syndrome are growth factors targeting either the granulocyte (Neupogen® or Neulasta®) or granulocyte and macrophage (Leukine®) hematopoietic cell lineages, there is interest in understanding the role that these factors play in responding to and/or ameliorating radiation damage. Furthermore, in an environment where resources are scarce, such as what might be expected during a radiation public health emergency, availability of growth factor or other treatments may be limited. For these reasons, the NIAID partnered with the Radiation Injury Treatment Network (RITN), whose membership includes medical centers with expertise in the management of bone marrow failure, to explore the use of growth factors and other cytokines as MCMs to mitigate/treat radiation injuries. A workshop was convened that included government, industry and academic subject matter experts, with presentations covering the anticipated concept of operations during a mass casualty incident including triage and treatment, growth factors under development for a radiation indication, and how the practice of medicine can inform other potential approaches, as well as considerations for administration of these products to diverse civilian populations. This report reviews the information presented, and provides an overview of the discussions from a guided breakout session.
Andrea DiCarlo; Zulmarie Horta; Jennifer Aldrich; Ann Jakubowski; William Skinner; Cullen Case


Use of Growth Factors and Cytokines to Treat Injuries Resulting from a Radiation Public Health Emergency.

Journal: Radiation research

Mai/7/2019
In response to concerns over possible radiological or nuclear incidents, the Radiation and Nuclear Countermeasures Program within the National Institute of Allergy and Infectious Diseases (NIAID) was tasked by the U.S. Department of Health and Human Services to support development of medical countermeasures (MCM) to treat the acute and delayed injuries that can result from radiation exposure. To date, the only three drugs approved by the U.S. Food and Drug Administration for treatment of acute radiation syndrome are growth factors targeting granulocyte (Neupogen® or Neulasta®) or granulocyte and macrophage (Leukine®) hematopoietic cell lineages. Although these are currently stockpiled for deployment in response to a mass casualty scenario, these growth factors will likely be administered in a scarce-resources environment and availability may be limited. Therefore, there is growing interest in understanding the role that these growth factors play in mitigating radiation damage, to optimize their use and maximize the number of people who can be treated. For these reasons, the NIAID and the Radiation Injury Treatment Network organized a workshop to explore the use of growth factors and other cytokines as MCMs in the treatment of radiation-induced injuries. Subject matter experts from government, industry and academia gathered at this workshop to discuss the concept of operations, triage and treatment, administration to diverse civilian populations, growth factors under development for radiation indications, and how the practice of medicine can inform other potential approaches.


Zulmarie Horta; Cullen Case; Andrea DiCarlo