Monoclonal Antibody 2–2-B Kills K1-positive Escherichia coli in Conjunction with Cord Blood Neutrophils and Sera, but Not with Spinal Fluid

Abstract Human blood neutrophils rolling on E- or P-selectin reduced their rolling velocity when intercellular adhesion molecule (ICAM)–1 was available. Similar to mouse neutrophils, this was dependent on P-selectin glycoprotein ligand 1 (PSGL1), αLβ2 integrin, the Src family tyrosine kinase FGR and spleen tyrosine kinase SYK. Blocking phospholipase C or p38 MAP kinase attenuated, but did not abolish the velocity reduction. To test expression of integrin activation epitopes, we adapted an immobilized reporter assay and developed a new homogeneous microfluidics-based reporter antibody binding assay. Rolling on E- or P-selectin induced the extension reporter epitopes KIM127 and NKI-L16, but not the high affinity reporter epitope monoclonal antibody (mAb) 24. This enabled rolling neutrophils to bind to immobilized extension reporter, but not activation reporter antibodies and allowed binding of soluble KIM127 during rolling. We conclude that human neutrophil rolling on E- or P-selectin induces the extended αLβ2 integrin conformation through signaling triggered by PSGL-1 engagement.

Download Full-text

F-actin content of neonate and adult neutrophils

Blood ◽

10.1182/blood.v69.3.945.bloodjournal693945 ◽

1987 ◽

Vol 69 (3) ◽

pp. 945-949 ◽

Cited By ~ 1

Author(s):

A Hilmo ◽

TH Howard

Keyword(s):

Cord Blood ◽

Gestational Age ◽

Actin Polymerization ◽

Flow Cytometric Analysis ◽

Relative Increase ◽

Chemotactic Response ◽

Similar Extent ◽

Cytoskeletal Organization ◽

Flow Cytometric ◽

Blood Neutrophils

We utilized flow cytometric analysis of NBDphallacidin-stained cells to measure F-actin content, expressed as fluorescent channel numbers, and we compared the microfilamentous cytoskeletal organization in neutrophils from healthy neonates (36 to 38 weeks gestational age) and adults. Basal F-actin content in neonate cord blood neutrophils is higher than that of adults. The elevation is intrinsic to the cell and not related to parturition because basal F-actin content of neonatal cells obtained by venipuncture (days 1 to 8 of age) is also elevated (38 +/- 10, N = 15) when compared to adults (21 +/- 7.0, N = 27). The rate of N-formyl-methionyl-leucyl-phenylalanine (fMLP)-induced actin polymerization is similar in adult and neonatal neutrophils and is maximal by 30 to 45 seconds at 25 degrees C. Adult, neonatal cord, and neonatal venipuncture neutrophils increase F-actin content to a similar extent following 0.5 mumol/L fMLP activation (52 +/- 18, N = 27; 58.7 +/- 18, N = 18; 51.5 +/- 7.0, N = 15, respectively). However, the relative increase in F-actin content following fMLP activation is much greater in adult (2.37-fold) than neonatal neutrophils (1.28-fold). This difference is due to the elevated basal F-actin content of neonatal cells. Comparison of distribution of F-actin content among basal, neonatal neutrophils reveals two subpopulations of neutrophils with respect to F-actin content--approximately 25% with F-actin content similar to that of adult neutrophils and 75% with F-actin content greater than that of adult cells. Following fMLP activation, the subpopulations disappear. The results suggest that abnormalities in microfilamentous cytoskeletal organization of neonatal cells may, in part, be responsible for decreased chemotactic response of neonatal neutrophils.

Download Full-text

Effect of Orally Administered Enterotoxigenic Escherichia coli K99-Specific Monoclonal Antibody to Neonatal Calves

Journal of the Japan Veterinary Medical Association ◽

10.12935/jvma1951.42.411 ◽

1989 ◽

Vol 42 (6) ◽

pp. 411-416

Author(s):

MASAO HORI ◽

SHUZO HIDAKA ◽

TSUGUO MIMURA

Keyword(s):

Escherichia Coli ◽

Monoclonal Antibody ◽

Enterotoxigenic Escherichia Coli ◽

Specific Monoclonal Antibody ◽

Neonatal Calves

Download Full-text

Protective Roles of γδ T Cells and Interleukin-15 in Escherichia coli Infection in Mice

Infection and Immunity ◽

10.1128/iai.66.7.3270-3278.1998 ◽

1998 ◽

Vol 66 (7) ◽

pp. 3270-3278 ◽

Cited By ~ 45

Author(s):

M. Takano ◽

H. Nishimura ◽

Y. Kimura ◽

Y. Mokuno ◽

J. Washizu ◽

...

Keyword(s):

Escherichia Coli ◽

Monoclonal Antibody ◽

T Cells ◽

T Cell Receptor ◽

Host Defense ◽

Γδ T Cells ◽

Cell Receptor ◽

E Coli ◽

Escherichia Coli Infection ◽

Interleukin 15

The number of γδ T cells in the peritoneal cavity was increased after an intraperitoneal (i.p.) infection with Escherichia coli in lipopolysaccharide (LPS)-responsive C3H/HeN mice but not in LPS-hyporesponsive C3H/HeJ mice. The γδ T cells preferentially expressed invariant Vγ6 and Vδ1 chains and proliferated to produce a large amount of gamma interferon in the presence of LPS. Mice depleted of γδ T cells by T-cell receptor δ gene mutation showed impaired resistance against E. coli as assessed by bacterial growth. Macrophages from C3H/HeN mice infected with E. coli expressed higher levels of interleukin-15 (IL-15) mRNA than those from the infected C3H/HeJ mice. Administration of anti-IL-15 monoclonal antibody inhibited, albeit partially, the appearance of γδ T cells in C3H/HeN mice after E. coli infection and diminished the host defense against the infection. These results suggest that LPS-stimulated γδ T cells play an important role in the host defense against E. coli infection and that IL-15 may be partly involved in the protection via an increase in the γδ T cells.

Download Full-text