Chemotactic activity and receptor binding of neutrophil attractant/activation protein-1 (NAP-1) and structurally related host defense cytokines: interaction of NAP-2 with the NAP-1 receptor

• Published in: Journal of leukocyte biology 1991-03, Vol.49 (3), p.258-265
• Main Authors: Leonard, Edward J., Yoshimura, Teizo, Rot, Antal, Noer, Kathleen, Walz, Alfred, Baggiolini, Marco, Walz, Daniel A., Goetzl, Edward J., Castor, C. William
• Format: Article
• Language: English
• Subjects: Analysis of the immune response. Humoral and cellular immunity; Biological and medical sciences; Chemotaxis, Leukocyte; Complement C5a - physiology; connective tissue activating peptide‐III; CTAP‐III; Cytokines - physiology; flow cytometry; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Humans; Immunobiology; In Vitro Techniques; Interleukin-8 - physiology; interleukin‐8; Ligands; Miscellaneous; Neutrophils - physiology; Peptide Fragments - physiology; Peptides - physiology; PF‐4; platelet; Platelet Factor 4 - physiology; platelet factor‐4; Receptors, Immunologic - physiology; Receptors, Interleukin-8A; Regulatory factors and their cellular receptors; Structure-Activity Relationship
• ISSN: 0741-5400; 1938-3673
• DOI: 10.1002/jlb.49.3.258

Neutrophil attractant/activation protein‐1 (NAP‐1) has sequence similarity to platelet factor‐4 (PF‐4) and to NAP‐2 (a truncated form of connective tissue activating protein‐Ill [CTAP‐III(des 1–15)]. We compared chemotactic activity for neutrophils of these related proteins. We also included for comparison CTAP‐III, CTAP‐III(des 1–13), the C‐terminal dodecapeptide of PF‐4 [PF‐4(59–70)], and C5a. Chemotactic potency (EC50) was highest for NAP‐1 and C5a. Although chemotactic efficacy (peak percentage of neutrophils migrating) was comparable for C5a, NAP‐1, and NAP‐2, the NAP‐2 response occurred only at concentrations 100‐fold higher than the NAP‐1 EC50 of 10‐8 M. Data for the CTAP‐III proteins confirmed that CTAP‐III is not an attractant and that chemotactic activity appears as a result of cleavage of residues at the N‐terminus to make CTAP‐III(des 1–13) or NAP‐2 [CTAP‐III(des 1–15)]. Chemotactic activity of PF‐4 was low and variable, with no significant response by neutrophils from six of nine subjects. In contrast, PF‐4(59–70) regularly induced high chemotactic responses, although the EC50 of 1.6 × 10‐5 M was 1,000‐fold greater than that of NAP‐1. The binding of fluoresceinated NAP‐1 to neutrophils was inhibited by unlabeled NAP‐1 or NAP‐2 but not by PF‐4 or PF‐4 (59–70). This suggests that NAP‐2 interacts with the neutrophil NAP‐1 receptor. Despite the low chemotactic potency of NAP‐2, it is a potential attractant at sites of injury because of the relatively large amounts of the parent CTAP‐III released from platelets, as indicated by a serum concentration of approximately 10‐6 M.