On Connective Tissue Mast Cells as Protectors of Life, Reproduction, and Progeny.

The connective tissue mast cell (MC), a sentinel tissue-residing secretory immune cell, has been preserved in all vertebrate classes since approximately 500 million years. No physiological role of the MC has yet been established. Considering the power of natural selection of cells during evolution, it is likely that the MCs exert essential yet unidentified life-promoting actions. All vertebrates feature a circulatory system, and the MCs interact readily with the vasculature. It is notable that embryonic MC progenitors are generated from endothelial cells. The MC hosts many surface receptors, enabling its activation via a vast variety of potentially harmful exogenous and endogenous molecules and via reproductive hormones in the female sex organs. Activated MCs release a unique composition of preformed and newly synthesized bioactive molecules, like heparin, histamine, serotonin, proteolytic enzymes, cytokines, chemokines, and growth factors. MCs play important roles in immune responses, tissue remodeling, cell proliferation, angiogenesis, inflammation, wound healing, tissue homeostasis, health, and reproduction. As recently suggested, MCs enable perpetuation of the vertebrates because of key effects-spanning generations-in ovulation and pregnancy, as in life-preserving activities in inflammation and wound healing from birth till reproductive age, thus creating a permanent life-sustaining loop. Here, we present recent advances that further indicate that the MC is a specific life-supporting and progeny-safeguarding cell.

Immunity to pathogenic fungi in the eye.

Fusarium, Aspergillus and Candida are important fungal pathogens that cause visual impairment and blindness in the USA and worldwide. This review will summarize the epidemiology and clinical features of corneal infections and discuss the immune and inflammatory responses that play an important role in clinical disease. In addition, we describe fungal virulence factors that are required for survival in infected corneas, and the activities of neutrophils in fungal killing, tissue damage and cytokine production.

Oropharyngeal tumor cells induce COX-2 expression in peripheral blood monocytes by secretion of IL-1α.

Oropharyngeal squamous cell cancer (OPC) accounts for 3% of all cancers and greater than 1.5% of all cancer deaths in the United States, with marked treatment-associated morbidity in survivors. More than 80% of OPC is caused by HPV16. Tumors induced by HPV have been linked to impaired immune functions, with most studies focused on the local tumor microenvironment. Fewer studies have characterized the effects of these tumors on systemic responses in OPC, especially innate responses that drive subsequent adaptive responses, potentially creating feed-back loops favorable to the tumor. Here we report that elevated plasma levels of PGE2 are expressed in half of patients with OPC secondary to overexpression of COX-2 by peripheral blood monocytes, and this expression is driven by IL-1α secreted by the tumors. Monocytes from patients are much more sensitive to the stimulation than monocytes from controls, suggesting the possibility of enhanced immune-modulating feed-back loops. Furthermore, control monocytes pre-exposed to PGE2 overexpress COX-2 in response to IL-1α, simulating responses made by monocytes from some OPC patients. Disrupting the PGE2/IL-1α feed-back loop can have potential impact on targeted medical therapies.

Analysis of Chitinase-3-Like Protein 1, IL-1-Alpha, and IL-6 as Novel Inflammatory Biomarkers for COVID-19.

The aim of our study was to investigate the potential role of IL-1-alpha, IL-6, and chitinase 3-like protein-1 (CHI3L1) as potential biomarkers for COVID-19. Sixty adult SARS Cov-2 PCR-positive patients (22 mild, 25 moderate, and 13 severe) and 50 healthy controls were included in this study. The serum levels of CHI3L1, IL-1-alpha, and IL-6 for all study participants were measured by protein-specific ELISAs. Mean serum CHI3L1 levels in patients with severe disease (7,185.5 ± 1,109.4) were significantly higher than in the moderate (3,977.4 ± 1,260.3), mild (1,379.5 ± 598.8), and control (329.5 ± 128.4) groups (P = 0.001). There was no difference in IL-1-alpha levels between the patient and control groups (P = 0.083). IL-6 levels differed significantly, being lowest in the control group (35.9 ± 13.7), 89.1 ± 23.4 in the mild group, 156.2 ± 29.6 in the moderate group, and the highest in the severe group (214.9 ± 28.1) (P = 0.001). A strong significant correlation was found between disease severity and serum IL-6 and CHI3L1 values (r = 0.894 and r = 0.905, respectively, and P < 0.001 for both). Serum CHI3L1 and IL-6 levels exhibited a linear correlation with the clinical course of COVID-19 infection. These results indicate that inhibitors of IL-6 and/or CHI3L1 may provide useful treatments for COVID-19.

Augmented secretion of IL-1α from mouse oral squamous cell carcinoma (OSCC) vcells caused by serum deprivation and hypoxia promotes immune suppressive activity of mesenchymal stromal cells.

OBJECTIVES: We have previously reported that mouse oral squamous carcinoma (OSCC), Sq-1979-1, produces interleukin (IL)-1α, which specifically enhances the immunosuppressive activity of co-cultured mesenchymal stromal 10T1/2 cells. This study assessed the conditions promoting the production of IL-1α in Sq-1979-1 cells, which could further enhance the immunosuppressive function of 10T1/2 cells, and evaluated its expression in OSCC tissues. METHODS: The expression of IL-1α was examined by RT-PCR, western blotting, and enzyme-linked immune sorbent assay (ELISA). The interferon (IFN)- γ-producing capability of anti-CD3 antibody-stimulated mouse spleen cells co-cultured with 10T1/2 cells and conditioned medium (CM) from Sq-1979-1 cells was examined by ELISA. The function of IL-1α was examined using an anti-IL1α antibody. Immunohistochemical analysis of the OSCC tissues was performed. RESULTS: The production of IL-1α from Sq-1979-1 cells was synergistically enhanced in lower serum (0.5% or 1.0% FBS) at the transcriptional level, and under hypoxia (1.0% oxygen) at the release level compared to that in the control medium supplemented with 10% FBS under normoxia. The IFN-γ-producing capability of stimulated spleen cells co-cultured with 10T1/2 cells was significantly reduced in the CMs prepared with the lower serum or under hypoxia. These functions of CMs were completely abolished by the anti-IL-1α antibody. The expression of IL-1α in OSCC tissues was prominent in the midst of a carcinomatous cellular lesion or a nearby necrotic lesion, where a supply deficiency could occur. CONCLUSION: s: IL-1α production by Sq-1979-1 cells was synergistically augmented under low serum and hypoxic conditions, which could promote the immunosuppressive activity of mesenchymal cells.

The Immunological Impact of IL-1 Family Cytokines on the Epidermal Barrier.

The skin barrier would not function without IL-1 family members, but their physiological role in the immunological aspects of skin barrier function are often overlooked. This review summarises the role of IL-1 family cytokines (IL-1α, IL-1ß, IL-1Ra, IL-18, IL-33, IL-36α, IL-36ß, IL-36γ, IL-36Ra, IL-37 and IL-38) in the skin. We focus on novel aspects of their interaction with commensals and pathogens, the important impact of proteases on cytokine activity, on healing responses and inflammation limiting mechanisms. We discuss IL-1 family cytokines in the context of IL-4/IL-13 and IL-23/IL-17 axis-driven diseases and highlight consequences of human loss/gain of function mutations in activating or inhibitory pathway molecules. This review highlights recent findings that emphasize the importance of IL-1 family cytokines in both physiological and pathological cutaneous inflammation and emergent translational therapeutics that are helping further elucidate these cytokines.

IL-1α cleavage by inflammatory caspases of the noncanonical inflammasome controls the senescence-associated secretory phenotype.

Interleukin-1 alpha (IL-1α) is a powerful cytokine that modulates immunity, and requires canonical cleavage by calpain for full activity. Mature IL-1α is produced after inflammasome activation and during cell senescence, but the protease cleaving IL-1α in these contexts is unknown. We show IL-1α is activated by caspase-5 or caspase-11 cleavage at a conserved site. Caspase-5 drives cleaved IL-1α release after human macrophage inflammasome activation, while IL-1α secretion from murine macrophages only requires caspase-11, with IL-1ß release needing caspase-11 and caspase-1. Importantly, senescent human cells require caspase-5 for the IL-1α-dependent senescence-associated secretory phenotype (SASP) in vitro, while senescent mouse hepatocytes need caspase-11 for the SASP-driven immune surveillance of senescent cells in vivo. Together, we identify IL-1α as a novel substrate of noncanonical inflammatory caspases and finally provide a mechanism for how IL-1α is activated during senescence. Thus, targeting caspase-5 may reduce inflammation and limit the deleterious effects of accumulated senescent cells during disease and Aging.

TNF-α and plasma albumin as biomarkers of disease activity in systemic lupus erythematosus.

OBJECTIVES: Composite criteria/indices are presently used to diagnose and monitor patients with systemic lupus erythematosus (SLE). Biomarkers for these purposes would be helpful in clinical practice. We therefore evaluated a large panel of cytokines and basic laboratory tests and investigated their performance as discriminators versus controls and as biomarkers of disease activity (DA). METHODS: We examined 437 patients with SLE, fulfilling American College of Rheumatology-82 criteria, and 322 matched controls. DA was assessed according to both SLE DA Index 2000 (SLEDAI-2K) and SLE Activity Measure (SLAM). British Isles Lupus Activity Group (BILAG) was used to assess renal DA. Additionally, 132 patients self-assessed their Global Disease Activity (PtGDA). Mesoscale Discovery 30-plex cytokine assay and routine blood chemistry was performed on fasting EDTA-plasma. RESULTS: Of 26 tested biomarkers, we identified TNF-α as the superior discriminator between patients with SLE and controls (median=4.5 pg/mL, IQR=3.1-6.2 vs median=2.3 pg/mL, IQR=2.0-2.8). The strongest correlations to SLEDAI-2K and SLAM were obtained with TNF-α (Spearman rho (ρ)=0.32 and ρ=0.34, respectively), partly driven by the nephritis subgroup, and with p-albumin (ρ=-0.33 and ρ=-0.31, respectively). P-albumin was decreased and TNF-α was increased in patients with kidney involvement (renal BILAG A/B vs C/D/E, p=4×10-16 and p=6×10-9 respectively). IP-10 was increased in patients with joint involvement (SLAM item 24≥2 vs ≤1, p=0.0005) but did not differ when comparing patients with active/inactive kidney involvement. The most powerful correlations to PtGDA was observed with p-albumin (ρ=-0.42), IL-6 (ρ=0.30) and TNF-α (ρ=0.29). CONCLUSION: TNF-α and p-albumin both performed well as discriminators between patients with SLE and controls and as proxies for DA according to both rheumatologists' and patients' assessments. In particular, renal DA was well reflected by TNF-α. We propose that the TNF-α and p-albumin merit further investigations as clinically useful biomarkers in SLE. We also observed that the pattern of activated cytokines varies with organ involvement.

Epinephrine stimulates CXCL1 IL-1α, IL-6 secretion in isolated mouse limb muscle.

Catecholamines stimulate interleukin-6 (IL-6) secretion in skeletal muscles. However, whether other cytokines are secreted is currently unknown. Skeletal muscle ex vivo preparations commonly used to study cytokine secretion have dealt with limitations including auto-oxidation of catecholamines. The use of metal chelators could be an alternative to avoid auto-oxidation and allow catecholamines to be used at physiological doses. We exposed isolated soleus muscles to 1 or 100 ng/mL epinephrine (EPI) and collected bath samples at 1 and 2 h for multiplex cytokine analysis. Keratinocyte chemoattractant (CXCL1), IL-6, and IL-1α were significantly elevated by 100 ng/mL exposure, but not by 1 ng/mL (median [CXCL1] (2 h) = 83 pg/mL; [IL-6] = 19 pg/mL; IL-1α = 7.5 pg/mL). CXCL1 and IL-6 were highly correlated in each sample (P = 0.0001). A second experiment combined the metal chelator, deferoxamine mesylate (DFO), to prevent EPI autoxidation, with 2 ng/mL EPI and 10.5 ng/mL norepinephrine (NOREPI) to mimic peak exercise. Unexpectedly, DFO alone stimulated both IL-6 and CXCL1 secretion, but together with EPI and NOREPI had no additional effects. Stimulation of cytokine secretory responses from skeletal muscle cells in response to DFO thus precludes its use as a chelating agent in ex vivo models. In conclusion, 100 ng/mL EPI stimulates a robust secretory CXCL1 response, which together with IL-6 and IL-1α, may constitute an adrenal-muscle endocrine response system.

Potential of IL-1, IL-18 and Inflammasome Inhibition for the Treatment of Inflammatory Skin Diseases.

In 2002, intracellular protein complexes known as the inflammasomes were discovered and were shown to have a crucial role in the sensing of intracellular pathogen- and danger-associated molecular patterns (PAMPs and DAMPs). Activation of the inflammasomes results in the processing and subsequent secretion of the pro-inflammatory cytokines IL-1ß and IL-18. Several autoinflammatory disorders such as cryopyrin-associated periodic syndromes and Familial Mediterranean Fever have been associated with mutations of genes encoding inflammasome components. Moreover, the importance of IL-1 has been reported for an increasing number of autoinflammatory skin diseases including but not limited to deficiency of IL-1 receptor antagonist, mevalonate kinase deficiency and PAPA syndrome. Recent findings have revealed that excessive IL-1 release induced by harmful stimuli likely contributes to the pathogenesis of common dermatological diseases such as acne vulgaris or seborrheic dermatitis. A key pathogenic feature of these diseases is IL-1ß-induced neutrophil recruitment to the skin. IL-1ß blockade may therefore represent a promising therapeutic approach. Several case reports and clinical trials have demonstrated the efficacy of IL-1 inhibition in the treatment of these skin disorders. Next to the recombinant IL-1 receptor antagonist (IL-1Ra) Anakinra and the soluble decoy Rilonacept, the anti-IL-1α monoclonal antibody MABp1 and anti-IL-1ß Canakinumab but also Gevokizumab, LY2189102 and P2D7KK, offer valid alternatives to target IL-1. Although less thoroughly investigated, an involvement of IL-18 in the development of cutaneous inflammatory disorders is also suspected. The present review describes the role of IL-1 in diseases with skin involvement and gives an overview of the relevant studies discussing the therapeutic potential of modulating the secretion and activity of IL-1 and IL-18 in such diseases.

Safety, pharmacokinetics, and preliminary efficacy of a specific anti-IL-1alpha therapeutic antibody (MABp1) in patients with type 2 diabetes mellitus.

AIMS: The role of the IL-1 system in development of type 2 diabetes is well established. Using an IL-1 receptor antagonist, which blocks IL-1alpha and -beta activity, or by specifically neutralizing IL-1beta, several clinical studies have demonstrated improvement in insulin secretion and glycaemia. However, the role of IL-1alpha remains to be investigated. METHODS: We evaluated the safety and preliminary efficacy of a neutralizing true human™ monoclonal antibody against IL-1alpha (MABp1) in an open label trial in patients with type 2 diabetes. Seven patients between 50 to 66years with type 2 diabetes mellitus were enrolled in the study. The study subjects received four biweekly intravenous infusions of MABp1 at 1.25mg/kg body weight up to day 60 and were followed up for a total of 90days. RESULTS: Compared to baseline, after the 60-day period of treatment HbA1c was numerically reduced by 0.14±0.21% (p=0.15), fasting C-peptide was increased by 88% (p=0.03), pro-insulin by 48% (p=0.03) and insulin numerically increased by 74% (p=0.11). Systolic blood pressure numerically decreased by 11mmHg (p=0.2). Both HbA1c and blood pressure rebounded to baseline levels thirty days after the end of MABp1 application. Treatment with MABp1 was well tolerated, and no adverse events occurred during the study. CONCLUSION: The results point to a role of IL-1alpha in type 2 diabetes and encourage further investigations. (ClinicalTrials.gov number NCT01427699).

IL-1 receptor antagonist improves morphine and buprenorphine efficacy in a rat neuropathic pain model.

An interesting research and therapeutic problem is the reduced beneficial efficacy of opioids in the treatment of neuropathic pain. The present study sought to investigate the potential role of IL-1 family members in this phenomenon. We studied the time course of changes in IL-1alpha, IL-1beta, IL-1 receptor type I and IL-1 receptor antagonist mRNA and protein levels experienced by rats after chronic constriction injury (CCI) of the sciatic nerve using qRT-PCR and Western blot analysis. In CCI-exposed rats, spinal levels of IL-1alpha mRNA were slightly downregulated on the 7th day, and protein levels were not changed on the 7th and 14th days. Levels of IL-1 receptor antagonist and IL-1 receptor type I were slightly upregulated in the ipsilateral part of the spinal cord on the 7th and 14th days; however, protein levels were not changed at those time points. Interestingly, we observed that IL-1beta mRNA and protein levels were strongly elevated in the ipsilateral part of the dorsal spinal cord on the 7th and 14th days following CCI. Moreover, in rats exposed to a single intrathecal administration of an IL-1 receptor antagonist (100 ng i.t.) on the 7th and 14th day following CCI, symptoms of neuropathic pain were attenuated, and the analgesic effects of morphine (2.5 µg i.t.) and buprenorphine (2.5 µg i.t.) were enhanced. In summary, restoration of the analgesic activity of morphine and buprenorphine by blockade of IL-1 signaling suggests that increased IL-1beta responses may account for the decreased analgesic efficacy of opioids observed in the treatment of neuropathy.

Differential expression of VEGF and IL-1alpha after photodynamic treatment in combination with doxorubicin or taxotere.

AIM: To show the impact of chemotherapeutic drugs doxorubicin and taxotere on the molecular pattern of cell response to photodynamic treatment (PDT). MATERIALS AND METHODS: Human squamous cell carcinoma cells A-431 were studied. Apoptosis was investigated by recording caspase-3 activity. Expression of IL-1alpha and VEGF on mRNA and protein levels was measured by qPCR and ELISA. RESULTS: PDT in combination with either doxorubicin or taxotere was found to be more cytotoxic in comparison to either single-treatment. The expression of IL-1alpha and VEGF was up-regulated in PDT-treated cells, either alone or in combination with doxorubicin or taxotere. Addition of doxorubicin to the cytokine induction after PDT was not detected, however, taxotere promoted significant over-expression of IL-1alpha and VEGF on the protein level. CONCLUSION: Contribution of chemotherapeutic drugs to IL-1 alpha and VEGF release from cells which received dual treatment involving PDT could be significantly different, despite the same level of cytotoxicity.

In Vitro Effects of 1,25-Dihydroxyvitamin D3 on the Production of Interleukin-1alpha by Ultraviolet B Irradiation in Cultured Human Keratinocyte Cell Line HaCaT Cells / 대한피부과학회지

BACKGROUND: Keratinocyte-derived interleukin-1(IL-1)alpha is one of the key cytokines in initiation of cutaneous inflammation. Release of IL-1alpha from human keratinocytes may be induced by proinflammatory stimuli including ultraviolet B(UVB) irradiation, and subsequently, keratinocyte-derived IL-1alpha may exert numerous paracrine and autocrine effects. 1,25-dihydroxyvitamin D3(1,25(OH)2D3) is involved in the regulation of keratinocyte proliferation and differentiation and is also recognized to have immunoregulatory properties such as an antiinflammatory effect. OBJECTIVE: The purpose of this study was to investigate the in vitro effects of 1,25-(OH)2D3 on the production of IL-1alpha by UVB irradiation in cultured human keratinocyte cell line HaCaT cells. RESULTS: are summerized as follows; 1. The vialility of cultured HaCaT cells measured by MTS assay at 24 hours after UVB irradiation was significantly reduced at the doses of above 100 mJ/cm2 of UVB(p<0.05). 2. The secretion of IL-1alpha by HaCaT cells was significantly increased at the doses of above 30 mJ/cm2 of UVB(p<0.05). UVB irradiation could not influence on the secretion of IL-1beta by HaCaT cells. 3. At the concentrations of 10-8M and 10-6M of 1,25(OH)2D3, the production of IL-1alpha by HaCaT cells(48 hours after 100 mJ/cm2 UVB irradiation) was significantly inhibited in both culture supernatants and cell lysates(p<0.05). CONCLUSION: UVB irradiation increased the production of IL-1alpha by HaCaT cells and this stimulatory effect on the production of IL-1alpha induced by UVB irradiation was suppressed by 1,25-(OH)2D3. Calcipotriol(MC-903) had similar suppressive effect on the production of IL-1alpha induced by UVB irradiation in HaCaT cells to that of 1,25(OH)2D3.

The Effects of CD11c/CD18 and CD14 Blocking on Lipopolysaccharide-induced Endotoxemia

We studied the morphologic changes and the expression of cytokines of major organs by blocking CD14 and CD11c/CD18, which are known to be receptors of lipopolysaccharide (LPS), in the LPS induced endotoxemic mice. In 2 and 8 hours after initial intraperitoneal injection of 10 mg/kg of LPS into the mice, 500 microgram/kg of anti-CD14 antibody (Ab) and/or the same dosage of the anti-CD11c/CD18 Ab were administered intravenously, respectively or concomitantly. Under the light microscope, the LPS only and the LPS with the anti-CD14 Ab injected groups (group 1 and 2) showed marked acute inflammation in the organs, especially in the lung and liver, but the LPS with the anti-CD11c/CD18 Ab only or with both anti-CD14 and anti-CD11c Abs injected groups (group 3 and 4) revealed only mild acute inflammation. Under the electron microscope, there was marked inflammation in the group 1 and 2 such as marked infiltration of neutrophils, monocytes/ macrophages, lymphocytes, aggregation of platelets, and marked edematous change of endothelial cells with separation from the basement membrane. However in the group 3 and 4, there was only mild inflammation such as mild infiltration of neutrophils in the tissue or aggregation of neutrophils in the capillaries and sinusoids with mild endothelial swelling. The expressions of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1alpha (IL-1alpha), detected by RT-PCR method, was remarkable in the group 2, but minimal in the group 3 and 4. We conclude that blocking the CD11c/CD18 with anti-CD11C/CD18 Ab is effective for the prohibition of biologic reactions and diminution of the inflammation induced by the LPS, even in the LPS induced endotoxemia.

Inhibitory Effects of Dexamethasone on Production IL-1alpha and TNF-alpha in Peripheral Blood Mononuclear Cells

PURPOSE: IL-1alpha and TNF-alpha have a seminal role in the intial events of inflammation. An area of intense interest is the assessment of new therapeutic modalities that regulate inflammatory response in acute bacterial infection. We conducted this study to compare the inhibitory effects of dexamethasone on the secretion of proinflammatory cytokines, such as IL-lalpha and TNF-alpha from cord blood mononuclear cells(MC) to those of adult blood MC during stimulation with S. aureus TSST-1 and E. coli LPS. METHODS: MC were isolated by differential centrifugation on Ficoll-Hypaque gradients. Each MC were incubated with TSST-1(2ug/ml) or LPS(0.2ug/ml), with various concentrations of dexamethasone for 72hr. And the other MC were incubated with TSST-1 or LPS, using the same concentration of dexamethasone, which was added 4hr before or simultaneously or 4hr, 24hr, 48hr after the stimulation. Concentration of IL-1alpha and TNF-alpha was measured by ELISA kit. RESULTS: Dexamethasone showed significant inhibitory effects on secretion of IL-1alpha and TNF-alpha. In comparison with both cytokines, secretion of IL-1alpha was suppressed more severely than TNF-alpha. In comparison with each stimulators, inhibition of TSST-1 induced cytokines production was greater than LPS. There was no difference between adult and cord blood MC. When dexamethasone was added to MC 4hr before the stimulation, it had the best inhibitory effects on the production of proinflammatory cytokines. CONCLUSION: Proinflammatory cytokine production in cord and adult blood MC were inhibited by dexamethasone in a dose-dependent manner. Early treatment of dexamethasone is more effective and can be used for modulating or suppressing excessive proinflammatory cytokine production in acute bacterial infection.

Inhibitory Effects of Actinomycin-D and Pentoxifylline on the Production of IL-1alpha and TNF-alpha by Human Cord and Adult Blood Mononuclear Cells

PURPOSE: An area of intense intrest is assessment of new therapeutic modalities that regulate the inflammatory response in acute bacterial infection is proving to be an area of interest these days. So we conducted this study to compare the inhibitory effects of actinomycin-D and pentoxifylline on the production of IL-lalpha and TNF-alpha from cord blood mononuclear cells (MC) to those from adult blood MC stimulated with S. aureus toxic shock syndrome toxin (TSST)-1 and E. coli lipopolysaccharide (LPS). METHODS: Cord and adult blood MC were isolated by differential centrifugation on Ficoll-Hypaque gradients. Each mononuclear cells were incubated with TSST-1 (2 microgram/ml) or LPS (0.2 microgram/ml), simultaneously with various concentrations of actinomycin-D or pentoxifylline added for inhibition. Concentration of interleukin-1alpha (IL-lalpha) and tumor necrosis factor-alpha (TNF-alpha) were measured by means of ELISA. RESULTS: In comparison with each inhibitory drug, actinomycin-D showed more potent inhibitory effects on the production of IL-1alpha and TNF-alpha from adult and cord blood MC stimulated by TSST-1 and LPS, than pentoxifylline (P<0.05). There was no difference between adult and cord blood MC. In comparison with each stimulator, inhibition of TSST-1 induced cytokines production with actinomycin-D was greater than pentoxifylline, in contrast to inhibition of LPS induced cytokines production with pentoxifylline which was greater than actinomycin-D in adult blood MC (P<0.05). CONCLUSION: Proinflammatory cytokine production in cord and adult blood MC were inhibited by each drug in the same manner except, the inhibition of pentoxifylline for LPS in cord blood MC. So it is possible that these drugs can be used to modulate or suppress excessive proinflammatory cytokine production in acute bacterial infection.

Production of Interleukin-1alpha and Tumor Necrosis Factor-alpha by Human Cord and Adult Blood Mononuclear Cells Stimulated by TSST-1 and LPS

PURPOSE: Immature immunological defens mechanism in the neonate may contribute to the high susceptibility to overwhelming sepsis. S. aureus TSST-1 and E. coli LPS known as one of the important pathogens of septic shock or toxic shock induce massive release of various cytokines, such as tumor necrosis factor-alpha (TNF-alpha) and interleukin-1alpha (IL-1alpha) produced from peripheral blood mononuclear cells (PBMC). In contrast, limited information has been provided so far concerning the capacity of cytokine production from neonatal immune cells. METHODS: This study was conducted to compare the secretion of proinflammatory cytokines, such as IL-l and TNF-alpha from cord blood PBMC to those from adult blood PBMC stimulated by S. aureus TSST-1 and E. coli LPS. RESULTS: 1) IL 1-alpha was secreted in a time-dependent manner from cord & adult blood PBMC stimulated with several cytokine inducers, and LPS stimulated adult & cord blood PBMC secreted IL 1-alpha in a dose-dependent manner. 2) TNF-alpha secretion from cord blood PBMC stimulated with LPS and IFN- significantly decreased in a time dependent manner, but not from adult PBMC. And secretion of TNF- from cord blood PBMC reached the highest level 24 hours after stimulated with LPS or IFN-gamma. The secretion of TNF-alpha from adult blood PBMC showed similar pattern to those from cord blood PBMC, but higher than cord blood PBMC. 3) IL-1alpha & TNF-alpha secretion from cord & adult blood PBMC stimulated with TSST-1 had no significant difference except in TNF- secretion by TSST-1 at 96 hours. 4) The secretion of IL-1alpha from adult PBMC stimulated with LPS showed higher and longer than that from cord blood PBMC. 5) IL-1alpha & TNF-alpha secretion from cord & adult blood PBMC stimulated with IFN-gamma had no significant difference. CONCLUSIONS: In summary, proinflammatory cytokines such as IL-1alpha, TNF-alpha in cord blood PBMC were secreted in a time dependent manner, but the amounts of IL-1alpha and TNF-alpha secretion were lesser than those of adult blood PBMC, especially stimulated by LPS. These results suggest that increased susceptibility to infection in neonatal period may be partially from a functional immaturity of cord blood mononuclear cells.