Novel recombinant human lactoferrin: Differential activation of oxidative stress related gene expression

Kruzel, Marian L.; Actor, Jeffrey K.; Zimecki, Michał; Wise, Jasen; Płoszaj, Paulina; Mirza, Shaper; Kruzel, Mark C.; Hwang, Shen An; Ba, Xueqing; Boldogh, István

doi:10.1016/j.jbiotec.2013.09.011

Cited by 58 publications

(58 citation statements)

References 57 publications

(70 reference statements)

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“…The methods for purification of the recombinant human LF have been published. 31 The level of endotoxin was <10 EU/mg protein. BCG M. bovis , Pasteur strain (TMC 1011; ATCC, Manassas, VA, USA) was grown in Dubos base (without addition of glycerol) with 10% supplement (5% BSA, 7.5% dextrose) on an orbital shaker at 37°C for 2 wk before use.…”

Section: Methodsmentioning

confidence: 97%

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Effects of CHO-expressed recombinant lactoferrins on mouse dendritic cell presentation and function

2014

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Lactoferrin (LF), a natural iron-binding protein, has previously demonstrated effectiveness in enhancing the Bacillus Calmette–Guérin (BCG) tuberculosis vaccine. This report investigates immune modulatory effects of Chinese hamster ovary (CHO) cell-expressed recombinant mouse and human LFs on mouse bone marrow-derived dendritic cells (BMDCs), comparing homologous and heterologous functions. BCG-infected BMDCs were cultured with LF, and examined for class II presentation molecule expression. Culturing of BCG-infected BMDCs with either LF decreased the class II molecule-expressing population. Mouse LF significantly increased the production of IL-12p40, IL-1β and IL-10, while human LF-treated BMDCs increased only IL-1β and IL-10. Overlaying naïve CD4 T-cells onto BCG-infected BMDCs cultured with mouse LF increased IFN-γ, whereas the human LF-exposed group increased IFN-γ and IL-17 from CD4 T cells. Overlay of naïve CD8 T cells onto BCG-infected BMDCs treated with mouse LF increased the production of IFN-γ and IL-17, while similar experiments using human LF only increased IL-17. This report is the first to examine mouse and human recombinant LFs in parallel experiments to assess murine DC function. These results detail the efficacy of the human LF counterpart used in a heterologous system to understand LF-mediated events that confer BCG efficacy against Mycobacterium tuberculosis challenge.

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Section: Methodsmentioning

confidence: 97%

“…31,32 The purity of the CHO-derived LFs was determined by HPLC and SDS PAGE, showing near homogeneity in protein (>98% purity). The methods for purification of the recombinant human LF have been published.…”

Section: Methodsmentioning

confidence: 99%

Effects of CHO-expressed recombinant lactoferrins on mouse dendritic cell presentation and function

2014

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“…Recombinant human and mouse LFs produced in CHO stable cell lines were provided by PharmaReview Corporation (Houston, TX) (22, 23). LFs were produced under endotoxin free serum conditions, and further affinity purified to remove traces of endotoxin (<0.1EU/mg in end product).…”

Section: Methodsmentioning

confidence: 99%

“…The bLF is only 70% homologous with human and mouse LFs (21), which raises concern on the applicability for use of heterologous LF in mouse or human model systems. Recently, we developed a CHO-based expression protocols for the production of both mouse and human recombinant LFs using a high expressing and stable cell line (DG44) (22). These novel recombinant LF proteins, with a mammalian type glycosylation patterns, can serve as useful tools to investigate LF in models of infection, while eliminating immunogenicity issues related to antigenic recognition of heterologous epitopes.…”

Section: Introductionmentioning

confidence: 99%

Immunomodulatory effects of recombinant lactoferrin during MRSA infection

Hwang¹,

Kruzel

Actor³

2014

International Immunopharmacology

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Methicillin-resistant Staphylococcus aureus (MRSA) infection remains a serious hazard to global health despite increases in public education and development of innovative treatment strategies. Use of immune modulatory therapy to combat infection is gaining interest as a novel treatment alternative. Lactoferrin (LF), an iron binding protein with multiple immune modulating properties, has the potential to modify the course of systemic MRSA infection. Specifically, LF is capable of limiting deleterious inflammatory responses while promoting development of antigen specific T-cell activity. The efficacy of a novel recombinant mouse LF (rmLF) to protect against MRSA infection was examined in a mouse peritonitis model. BALB/c mice were infected with a lethal dose of MRSA and treated at 2 hours post-infection with rmLF. The effects of rmLF on MRSA-infected primary monocytes and granulocytes were analyzed for inflammatory mediator production. The rmLF treated mice demonstrated only modest increase in survival by more than 24hrs, albeit with reduced bacteremia. Serum cytokines, IL-17 and IL-6, were significantly reduced post challenge in the rmLF treated mice. Treatment with rmLF led to a minor decrease in IL-1β, and a slight increase in TNF-α production. Preliminary investigation towards human clinical relevance was accomplished using human blood derived monocytes and granulocytes infected with MRSA and treated with a homologous recombinant human LF (rhLF). Treatment with (rhLF) led to increased production of IFN-γ and IL-2. The human cell studies also showed a concurrent decrease in TNF-α, IL-6, IL-1β, IL-12p40, and IL-10. The study reports the first investigation into the efficacy of a novel recombinant mouse lactoferrin (LF) therapy in a mouse model of MRSA peritonitis. Overall, these results indicate the rmLF and rhLF have a high degree of overlap to modify inflammatory responses, although differences in activities were observed indicating existence of mechanisms between the two heterologous recombinant molecules.

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“…The transition metal captors associate to metals known as pro-oxidants, such as copper and iron [57,58]. The main representatives of this class are the transferrin [59], lactoferrin [60], ferritin [61], albumin and ceruloplasmin [62]. However, their concentration is not dependant on the quantity of circulating free radicals, that is, an individual under constant physical activity will have greater production of ROS without the proportional elevation of lactoferrin for example.…”

Section: Physical Exercise Antioxidants and Oxidative Stressmentioning

confidence: 99%

Oriented Physical Exercise and Sports Nutrition and the Risk of Growth of Tumours

Tratz¹

2014

CRJ

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Abstract:The health and life quality of men can be preserved and improved using a regular practice of physical activities. It is well known that when properly oriented, the physical exercise can combat several diseases, e. g. cardiovascular, metabolic and even physiological, such as depression. However, the higher oxygen consumption, as well as the activation of specific metabolic pathways during and after the exercises results in the formation of free radicals. Exhausting physical exercises of high intensity elevate the risk of diseases, like pulmonary emphysemas, inflammation, aging and increased risk of damage in the DNA with consequent carcinogenesis. The excessive elevation of free radicals without the optimal accompaniment of antioxidants induces a psychopathological stage known as oxidative stress, which generates lipids, damages to proteins and to the DNA, becoming cytotoxic or causing changes in the cellular functions and even in the tissues. This is a common occurrence in drug users, overexposure to environmental factors such as sun light, metabolic alterations originated from obesity or diabetes, and even physical exercises. There is general agreement that physical training and adequate alimentation, besides protecting against some types of cancer, increases the aspects of anti-tumor immunity and reduces inflammatory mediators. Evidences suggest that the prophylactic effect of the exercise can be attributed to the anti-inflammatory effects mediated by the regular practice of exercises, through the reduction of visceral fat and increase of anti-inflammatory cytokines in the circulation. This study had as objective to demonstrate the importance of a well targeted training, for the performance of physical activities in an adequate intensity associated to antioxidant supplementation, in order to prevent excessive physical wear and appearing of oxidative stress.

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Novel recombinant human lactoferrin: Differential activation of oxidative stress related gene expression

Cited by 58 publications

References 57 publications

Effects of CHO-expressed recombinant lactoferrins on mouse dendritic cell presentation and function

Effects of CHO-expressed recombinant lactoferrins on mouse dendritic cell presentation and function

Immunomodulatory effects of recombinant lactoferrin during MRSA infection

Oriented Physical Exercise and Sports Nutrition and the Risk of Growth of Tumours

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