Studies of Chemotactic, Chemotactic Movement-Inhibiting and Random Movement-Inhibiting Effects of Interleukin-1 Alpha and Beta, Tumor Necrosis Factors Alpha and Beta and Interferon Gamma on Human Neutrophils in Assays Using ‘Sparse-Pore’ Polycarbonate (Nuclepore) Membranes in the Boyden Chamber

Previous studies have suggested that, in vivo, activated T lymphocytes and neutrophils are important in immunity to nontypable Haemophilus influenzae. We now extend this work by showing that neutrophils pretreated with products of activated T lymphocytes or activated macrophages show significantly enhanced killing of nontypable H. influenrae. Lymphotoxin, a product of activated T lymphocytes, significantly enhanced the neutrophil-mediated killing of nontypable H. infi uenzae, and tumor necrosis factor, produced by activated T lymphocytes as well as macrophages stimulated by activated T lymphocytes, also significantly increased the bactericidal activity of neutrophils. These cytokine-induced effects were seen with short pretreatment times of neutrophils and were maximal by 30 min. The killing of H . infiuenzae by neutrophils required the presence of heat-labile opsonins. In the absence of these opsonins, both tumor necrosis Nontypable Haemoph ilus infiuenzae is a co mmo n resp iratory pa thogen. It is carried in the nasopharynx of 70-80% of health y childr en (1). Coloniza tio n rates incr ease dur ing respirato ry illn ess (2) . In particul ar, colonizat ion of the nasoph arynx w ith nontypable H. infi uenzae may exceed 95% at the time of otitis media (3) , when the organism may compose 50% of the total bacterial flora (4). Although invasive disease, such as bacteremia, is uncommon with nontypabl e H. infl uenza e, local infection is frequent. Nont ypable H. infiu enzae is the second leading cause of acute otitis media and sinusitis and the lead ing ca use of recurrent or chronic otitis media in children (5-8).Th e protective role of antibody against nont ypable H. infiuenzae respi ratory infections in both humans and rat mod els is now qu estion abl e. High levels of antibo dy to this org an ism are presen t in se rum, saliva, and sputum of patients with chronic bronchitis (9, 10) , and in on e study the level of anti body in

show abstract

“…These include LT (28 -31), interferon-y (also a major product of natural killer cells) (29)(30)(31)(32)(33)(34), and IL-2 (35,36).…”

mentioning

confidence: 99%

“…These include TNF (30,33,(37)(38)(39)(40)41), IL-l (27,29), granulocy te-colonystimulating factor (34,42), and IL-8 (43).…”

mentioning

confidence: 99%

Activation of the Neutrophil Bactericidal Activity for Nontypable Haemophilus influenzae by Tumor Necrosis Factor and Lymphotoxin

et al. 1995

Self Cite

View full text Add to dashboard Cite

show abstract

“…Migration assays were performed using polycarbonate membrane chambers (pore size, 8.0 µm; EMD Millipore, Billerica, MA, USA) as previously demonstrated (27). Briefly, NCI-H1993 cells (1x10 5 ) were suspended in 100 µl RPMI-1640 medium containing 1.5% FBS were added to the upper chambers.…”

Section: Methodsmentioning

confidence: 99%

Synergistic antitumor activity of low-dose c-Met tyrosine kinase inhibitor and sorafenib on human non-small cell lung cancer cells

Guo

Zheng

et al. 2018

Oncol Lett

View full text Add to dashboard Cite

“…No. 111106) polycarbonate membranes were supplied by Nucleporc Corp (Calif.) and used after removal of wetting agent as previ ously described [24,27,28,30].…”

Section: Polycarbonate Membranesmentioning

confidence: 99%

“…First, adhesion and spreading can be assessed on the same mem brane [27] and second, studies of random motility can be carried out using polycarbonate membrane of standard pore density in chambers similar to that used in the Che motaxis assay [27,28]. In using the one type of substratum throughout the various assays, the problem of diverse properties of different substrata in these different assays is avoided.…”

Section: Introductionmentioning

confidence: 99%

lnterleukin-8 and Neutrophil Leucocytes: Adhesion, Spreading, Polarisation, Random Motility, Chemotaxis and Deactivation in Assays Using ‘Sparse-Pore’ Polycarbonate (Nuclepore) Membranes in the Boyden Chamber

Bignold

Harkin

Rogers³

1992

Int Arch Allergy Immunol

Self Cite

View full text Add to dashboard Cite

Adhesion, spreading, chemotaxis and deactivation of chemotactic responses of separated human peripheral blood neutrophil leucocytes under the influence of interleukin-8 (IL-8) were tested using a previously described Boyden chamber-type method involving ‘sparse-pore’ polycarbonate (Nuclepore) filtration membrane. The random motilities of neutrophils in similar concentrations of IL·8 were tested using corresponding chambers with polycarbonate membranes of standard pore densities. In addition, polarisation of neutrophils in suspension in various concentrations of IL-8, and the possibility of deactivation of this polarisation response by IL-8 itself or by N-formyl-methionyl-leucyl-phenylalanine (FMLP) were examined. Neutrophils exhibited maximum chemotaxis to IL-8 in a concentration of 100 ng/ml, and to a degree which was similar to the maximum response to FMLP (10^––⁷M). This chemotactic response to IL-8 was markedly reduced by pretreatment of the cells with either 100 ng/ml IL-8 (deactivation) or 10^––7M FMLP (cross-deactivation). On the other hand, the chemotactic response of the neutrophils to FMLP was reduced by pretreatment with FMLP but was not deactivated by pretreatment with IL-8 (i.e. deactivation but not cross-deactivation). Neutrophils in suspension were maximally polarised by 100 ng/ml IL-8, and to lesser degrees by 1,10 and 1,000 ng/ml IL-8. The detailed morphology of the polar cells was not distinguishable from that induced by 10^––8M FMLP. Pretreatment of the cells with either IL-8 or FMLP resulted in no reduction of polarisation in response to subsequent exposure to either agent (i.e. neither deactivation nor cross-deactivation). No effects of IL-8 on adhesion, spreading or random motility of neutrophils on the polycarbonate membranes were detected.

show abstract

Studies of Chemotactic, Chemotactic Movement-Inhibiting and Random Movement-Inhibiting Effects of Interleukin-1 Alpha and Beta, Tumor Necrosis Factors Alpha and Beta and Interferon Gamma on Human Neutrophils in Assays Using ‘Sparse-Pore’ Polycarbonate (Nuclepore) Membranes in the Boyden Chamber

Cited by 18 publications

References 19 publications

Activation of the Neutrophil Bactericidal Activity for Nontypable Haemophilus influenzae by Tumor Necrosis Factor and Lymphotoxin

Activation of the Neutrophil Bactericidal Activity for Nontypable Haemophilus influenzae by Tumor Necrosis Factor and Lymphotoxin

Synergistic antitumor activity of low-dose c-Met tyrosine kinase inhibitor and sorafenib on human non-small cell lung cancer cells

lnterleukin-8 and Neutrophil Leucocytes: Adhesion, Spreading, Polarisation, Random Motility, Chemotaxis and Deactivation in Assays Using ‘Sparse-Pore’ Polycarbonate (Nuclepore) Membranes in the Boyden Chamber

Contact Info

Product

Resources

About