Lactobacillus plantarum inhibits the intestinal epithelial migration of neutrophils induced by enteropathogenic Escherichia coli

BACKGROUND: Lactobacillus plantarum is a Gram-positive bacillus known for its effect as a probiotic agent. The goal of the study was to determine whether L. plantarum is capable of inhibiting the transepithelial neutrophil migration induced by enteropathogenic Escherichia coli (EPEC). METHODS: Cultured intestinal epithelial T-84 cell monolayers were rapidly infected with EPEC. L. plantarum or culture supernatants were added to the monolayers before and after the infection. Indium-labeled neutrophils were added to the basolateral side of inverted monolayers. After 150-minute incubation, radioactivity of the neutrophils that migrated in the physiologic direction was assayed, and the number of migrating neutrophils was calculated. L. plantarum was also added to the monolayers before and after EPEC infection, and the number of adherent EPEC was determined by plate counting. RESULTS: EPEC-induced neutrophil migration and EPEC binding to monolayers were inhibited by viable L. plantarum but only when added to the monolayers before EPEC. Culture supernatants failed to inhibit the neutrophil migration. CONCLUSIONS: These results suggest that L. plantarum is beneficial in inhibiting neutrophil migration induced by EPEC, but only when preincubated with host epithelia. Rather than an indirect effect through a secreted substance produced by the probiotic agent, its effect is direct and requires the presence of the bacterium.     

Effect of natural porcine surfactant (Curosurf) on the function of neutrophilic granulocytes

In this study we have analyzed various phagocytic functions of human neutrophils exposed to either biochemically well defined porcine surfactant (Curosurf) or a phospholipid preparation. Adherence, random migration and chemotactic response to zymosan activated serum and formyl-methionyl-leucyl-phenylalanine were normal in surfactant treated neutrophils; surfactant was not a chemotactic stimulus. In contrast, phagocytosis of S. aureus by neutrophils exposed to surfactant (100 micrograms/ml) or phospholipids (100 micrograms/ml) was impaired (surfactant: t30 49.5 +/- 9.0%, t60 65.0 +/- 8.0%; phospholipids: t30 66.3 +/- 12.6%, t60 78.0 +/- 7.8%; controls: t30 78.1 +/- 8.9%, t60 90.1 +/- 6.2%; p less than 0.001 at t30, t60 for surfactant, p less than 0.05 at t60 for phospholipids). Due to the smaller number of S. aureus ingested, bactericidal activity of surfactant or phospholipid treated neutrophils was slightly reduced when compared to controls (surfactant t30 p less than 0.05, surfactant t60 p less than 0.001, phospholipids t60 p less than 0.05). Surfactant or phospholipids had no bactericidal activity. Uptake of candida was identical in surfactant or phospholipid treated neutrophils with untreated controls; the same was true with the number of candida per cell ingested. Phagocytosis-associated chemiluminescence and production of superoxide anion by neutrophils of either source in response to phorbol myristate acetate and opsonized zymosan was also identical.     

A new model for studying eosinophil migration across cultured intestinal epithelial monolayers

OBJECTIVE: Eosinophils play an important role in some gastrointestinal inflammatory conditions. Stimulated eosinophils migrate across the vascular endothelial wall and into the intestinal epithelium where by-products such as proteases may contribute to intestinal epithelial damage. Little is known about the epithelial migration of the eosinophils in the gut. The lack of data is attributable in part to the scarcity of human eosinophils for studies. HL-60-differentiated eosinophils present a means to perform studies on eosinophil function and chemotaxis. HL-60 clone 15 can be induced to differentiate into cells closely resembling human eosinophils. The authors describe a novel model for studying eosinophil migration across the intestinal epithelium. METHODS: Fluorescent-labeled HL-60 eosinophils were incubated for 150 minutes on the basolateral surface of confluent and inverted T-84 monolayers separated by fluoroblock insert membranes. Chemotactic gradients of n-formyl methionyl leucyl phenylalanine (fMLP), eotaxin, and platelet aggregating factor (PAF) were used in variable concentrations. Changes in transepithelial electrical resistance (TEER) were compared with baseline values. RESULTS: Differentiated HL-60 eosinophils undergo migration in response to fMLP, PAF, and eotaxin. Migration is associated with a drop in TEER. CONCLUSION: In this model, HL-60-differentiated eosinophils migrate in response to stimulants chemotactic for human eosinophils. The transepithelial migration of eosinophils is associated with epithelial barrier dysfunction, which may contribute to the development of disease.     

The characterization of superoxide production of human neonatal neutrophil

To assess the role of neutrophil in neonatal host defense against microbial infection, we characterized the superoxide anion (O(2-)) production of neonatal neutrophil on a biochemical basis. After taking an appropriate informed consent, neutrophils were obtained from cord blood immediately after transvaginal delivery and divided into two groups: the Preterm group, 15 neonates (27-36 weeks' gestation) and the Term group, 15 neonates (37-41 weeks' gestation). Eleven healthy adults served as controls in the Adult group. The value of N-formyl-methionyl-leucyl-phenylalanin (fMLP)-induced O(2-) production activity of neutrophils in the Preterm group using chemiluminescence assay was significantly lower than those values in both the Term and Adult groups (5.77+/-0.53x10(6) vs. 11.1+/-0.94x10(6) and 10.7+/-0.63x10(6) cpm; mean+/-S.E.M., p<0.05). In phorbol 12-myristate 13-acetate (PMA)-stimulation, the values of O(2-) production activity of neutrophils in both the Preterm and Term groups were significantly lower than that in the Adult group (13.0+/-1.66x10(6) and 18.0+/-1.44x10(6) vs. 27.3+/-1.45x10(6) cpm, p<0.05). Scatchard analysis of [(3)H]fMLP binding to neutrophil demonstrated a two-receptor model in each group, and the number of high-affinity receptors per neutrophil in the Preterm group was significantly lower than those in other groups (p<0.05). However, cord blood levels of proinflammatory cytokines, such as interleukin (IL)-6, -8, and tumor necrosis factor-alpha (TNF-alpha) did not differ in either neonatal group. These results indicated that the fMLP-induced O(2-)production activity of neutrophils in the term neonates was enhanced at the level of the receptor and suggested that this enhanced production contribute to the neonatal host defense against microbial infection.     

Effect of granulocyte and granulocyte macrophage colony stimulating factors (G-CSF and GM-CSF) on neonatal neutrophil functions

Although there are many studies on the effect of granulocyte and granulocyte-macrophage colony stimulating factors (G/GM-CSF) on adult neutrophil functions, there is little information regarding their influence on neonatal cells. We studied the in vitro effect of G/GM-CSF on neutrophil chemotaxis, polarization, and superoxide anion generation in 47 neonates compared with 35 adults. We found that G-CSF and GM-CSF significantly enhanced the chemotaxis of newborn infants' neutrophils, normalizing their chemotactic defect [from 35 +/- 7 cells/field (mean +/- SE) to 49 +/- 5 cells/field with G-CSF, p < 0.05 and to 55 +/- 4 cells/field with GM-CSF, p < 0.001]. It is notable that the maximal neutrophil response to the cytokines was observed particularly in the newborn infants with severe impairment in their chemotactic activity. Statistical analysis of the data showed a significant inverse correlation, which supported this observation (r = -0.6, p < 0.02 for G-CSF; r = -0.76, p < 0.001 for GM-CSF). The reduced polarization of neonatal compared with adult cells [71 +/- 5% versus 86 +/- 2% (mean +/- SE), p < 0.05], was corrected by CSF-priming (to 87 +/- 4% with G-CSF and to 92 +/- 2% with GM-CSF, p < 0.05). In addition, the neutrophil superoxide generation was significantly improved in both groups following the CSF-priming. GM-CSF and G-CSF gave comparable results in all functions studied except that GM-CSF improved superoxide release to a greater extent. This study shows a significant improvement of the neonatal neutrophil functions following in vitro CSF-priming and contributes to a better understanding of the neonatal neutrophil behavior when treated with G/GM-CSF.     

Ontogeny of neutrophil chemotaxis in fetal lambs

Ontogeny of neutrophil chemotactic response using endotoxin activated adult sheep plasma as a source of complement derived chemotactic factor was examined in fetal lambs of gestational age 120-150 days. (Gestational period in sheep is approximately 150 days.) Neutrophils from fetal lambs of gestational age 120-130 days failed to respond to this chemotactic factor whereas neutrophils from fetal lambs above 131 days of gestational age responded at levels comparable to adult values. Examination of neutrophil chemotaxis in older fetuses using a different chemotactic factor derived from mitogen stimulated adult mononuclear cells revealed a selective failure of fetal neutrophils to respond to lymphocyte-derived chemotactic factor in the presence of a normal response to complement derived chemotactic factor. Among prematurely delivered twin fetuses alterations in comparison with the first twin or age-matched controls in peripheral neutrophil count (increase) and in chemotaxis (increase or decrease) were noted in second of twins delivered greater than or equal to 20 min after the first lamb, suggesting an extreme sensitivity of neutrophil functions to a variety of influences, similar to that seen in humans.     

Effects of neutrophil migration inhibitory factors on neonatal neutrophils

Neonatal neutrophil migration was inhibited by preincubation with a lymphokine/monokine-rich medium conditioned by phytohemagglutinin-stimulated mononuclear leucocytes. Medium conditioned by unstimulated mononuclear cells or nonconditioned medium had no effect on neonatal neutrophil migration. Similar results were obtained with adult neutrophils. Migration distances in the presence and absence of a chemotactic gradient were much lower for neutrophils from neonates than adults when comparing treatments with the corresponding medium, i.e. medium conditioned by phytohemagglutinin-stimulated mononuclear leucocytes, medium conditioned by unstimulated mononuclear leucocytes, or medium unconditioned by mononuclear leucocytes (p less than 0.01). Although locomotion of both neonatal and adult neutrophils was inhibited by treatment, the percent inhibition of random migration was slightly but significantly less for neonates than adults (p less than 0.05). These results demonstrate that neutrophils from neonates are modulated by mononuclear leucocyte-derived mediators.     

The effect of hybridoma antibody administration upon neutrophil kinetics during experimental type III group B streptococcal sepsis

Groups of newborn rats were transthoracically inoculated with 1 X 10(6) type III group B streptococci/g body wt, either alone or in combination with 1.5 microgram/g body wt of type-specific antibody derived from hybridoma cell lines. Ninety-four percent of the animals who received bacteria alone died. In contrast, none of those treated with antibody died (P less than 0.005). Kinetic studies suggested that antibody may have offered protection, In part, by facilitating the neutrophil response. Animals who received only bacteria exhibited a marked neutropenia (20 +/- 18/mm3, mean +/- S.E.M.) whereas infected animals treated with antibody did not (3800 +/- 30/mm3, P less than 0.001). Furthermore, within 2 h of inoculation, antibody-treated animals mobilized and stored neutrophils, whereas significant neutrophil mobilization did not occur in the animals which received bacteria alone until 6 h. In the animals receiving bacteria alone, exhaustion of the neutrophil supply quickly occurred (remaining storage neutrophils at 6 h, 0.2 +/- 0.1 X 10(6) cells). In contrast, animals, which received antibody, maintained an adequate supply of stored neutrophils (7.0 +/- 0.4 X 10(6) P less than 0.001). The migration of neutrophils to the site of inoculation was measured by assaying the lungs' content of myeloperoxidase, a marker enzyme for granulocytes. The right and left lungs of animals not receiving antibody accumulated the same quantity of neutrophils, with peak pulmonary neutrophil accumulation occurring 6 h after the infection. In antibody recipients, however, the inoculated lung accumulated significantly more neutrophils than the opposite lung and peak pulmonary neutrophil accumulation occurred at 2 rather than 6 h.     

Decreased adherence, chemotaxis and phagocytic activities of neutrophils from preterm neonates

Using microanalytic assays various phagocytic functions of separated neutrophils from preterm neonates (mean birthweight 1,506 g, n = 13) were simultaneously studied. Adherence of neutrophils to nylon fibre was decreased in cells from preterm infants (77.1 +/- 3.1%) when compared with adult controls (86.9 +/- 2.1%, mean +/- 1 SD, p less than 0.05). In addition neutrophil chemotaxis in response to zymosan activated serum was reduced in preterm neonates (131.9 +/- 19.7, adults 166.6 +/- 11.1, p less than 0.001); directed migration towards Formyl-Methionyl-Leucyl-Phenylalanine was also decreased (preterm neonates 93.4 +/- 15, adults 111.1 +/- 16.8, p less than 0.05). Preterm infants had a higher percentage of slow moving neutrophils when compared with adults (p less than 0.001). Phagocytosis of Candida albicans was reduced in neutrophils from preterm neonates (phagocytic index: preterm neonates 41.4 +/- 12.7, adults 83 +/- 7.2). Adult neutrophils ingested more Candida per cell (p less than 0.001). Chemiluminescence, exocytosis of elastase and lactoferrin during uptake of opsonized zymosan was also reduced in neutrophils from preterm neonates. However, random migration, phagocytosis of Staphylococcus aureus and production of O2- in response to Phorbol myristate acetate or opsonized zymosan were identical in cells from either source. We conclude, that these abnormalities of neutrophils could predispose the preterm infant to serious and often overwhelming bacterial and fungal infections.     

Susceptibility of human and porcine neutrophils to hypothermia in vitro

Hypothermia may contribute to serious life-threatening infections. An experimental model has been established in pigs in order to study the effects of hypothermia on host bacterial defenses. The function of blood neutrophils from pigs and humans was examined in vitro at 37 and 29 degrees C. Bacterial killing of Staphylococcus aureus 502A by human neutrophils after 90 and 180 min incubation at 29 degrees C was reduced to 76 +/- 6% and 83 +/- 7% of killing at 37 degrees C. Porcine neutrophil killing was similarly reduced at 90 min (72 +/- 9%) and remained significantly impaired after 180 min (52 +/- 11%). Phagocytosis of ORO-DP-LPS particles by human neutrophils after 5 min at 29 degrees C was 40 +/- 5% of that at 37 degrees C and only 55 +/- 7% after 15 min by which time maximum phagocytosis had occurred at 37 degrees C. Porcine neutrophils ingested significantly less ORO (68 +/- 8%) after 5 min at 29 degrees C and reached normal values by 15 min. Stimulation of hexose monophosphate pathway in human neutrophils for 20 min at 29 degrees C was only 13 +/- 5% of that at 37 degrees C and required 2 h of stimulation to reach normal values. Porcine cells were reduced to 74 +/- 9% after 20 min incubation and reached normal values by 30 min. Directed neutrophil migration as assessed under agarose was impaired for both human (39 +/- 6%) and porcine (20 +/- 4%) neutrophils at 29 degrees C compared to 37 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)     

Defective neutrophil motility in hypovitaminosis D rickets.

The chemotactic activity and random motility of neutrophils, was studied in 38 patients with hypovitaminosis D rickets, and compared with 29 healthy controls of matched age. The chemotactic activity derived from the activated rickets serum as well as the amounts of the complement components C4, C3 and C5 was normal, but the cell motility was clearly defective (p less than 0.001). A possible relationship between defective neutrophil movement and the recurrent infections seen in these patients is suggested. The possible mechanisms responsible for the defect could be the alteration in Ca/P metabolism or a defective action of the vitamin D on the neutrophils.     

Superoxide-dependent chemotactic activity for PMNs derived from opsonized zymosan-stimulated human platelets

Previous studies have shown that platelets exhibit a H2O2 producing, NADH-dependent system that is activated by interaction with particulate material. Current evidence suggests that this system could be critically involved in th generation of chemotactic factor(s). In the present studies, chemotactic activity for polymorphonuclear leukocytes of supernatants derived from zymosan-stimulated human platelets has been evaluated using an agarose gel technique. Supernatants of opsonized zymosan-stimulated platelets showed significant chemotactic activity (migration index = 300 +/- 50), in comparison with supernatants prepared from platelet suspensions stimulated with nonopsonized-zymosan (migration index = 10 +/- 15) or resting platelet supernatants (migration index = 15 +/- 15). Furthermore, a marked increase in chemotactic activity of the opsonized zymosan-treated platelet supernatants was demonstrated after the addition of NADH (migration index = 525 +/- 100). The inclusion of specific inhibitors of the cycloxygenase and lipoxygenase pathways resulted in a marked reduction of chemotactic activity, which was restored in the presence of NADH. Further, the addition of superoxide dismutase completely abolished the chemotactic response induced by NADH. These data suggest that platelets are the source of chemotactic factor(s) derived from the activation of a superoxide generating system.     

DEFECTIVE NEUTROPHIL MOTILITY IN HYPOVITAMINOSIS D RICKETS

ABSTRACT. The chemotactic activity and random motility of neutrophils, was studied in 38 patients with hypovitaminosis D rickets, and compared with 29 healthy controls of matched age. The chemotactic activity derived from the activated rickets serum as well as the amounts of the complement components C4, C3 and C5 was normal, but the cell motility was clearly defective (p<0.001). A possible relationship between defective neutrophil movement and the recurrent infections seen in these patients is suggested. The possible mechanisms responsible for the defect could be the alteration in Ca/P metabolism or a defective action of the vitamin D on the neutrophils.     

DEFECTIVE NEUTROPHIL MOTILITY IN HYPOVITAMINOSIS D RICKETS

Abstract. The chemotactic activity and random motility of neutrophils, was studied in 38 patients with hypovitaminosis D rickets, and compared with 29 healthy controls of matched age. The chemotactic activity derived from the activated rickets serum as well as the amounts of the complement components C4, C3 and CS was normal, but the cell motility was clearly defective ( p <0.001). A possible relationship between defective neutrophil movement and the recurrent infections seen in these patients is suggested. The possible mechanisms responsible for the defect could be the alteration in Ca/P metabolism or a defective action of the vitamin D on the neutrophils.     

Lactobacillus plantarum reduces the in vitro secretory response of intestinal epithelial cells to enteropathogenic Escherichia coli infection

OBJECTIVE: Enteropathogenic (EPEC) is a Gram-negative bacillus that causes diarrhea. Secretory responses of intestinal epithelial cells can be seen after EPEC infection. Probiotics, which are live bacteria that have proven benefit to the host, play a role in the treatment and prevention of the different enteric pathogens. The goals of the study were to determine whether the probiotic agent (LBP) strain 299v alters the secretory changes seen in EPEC infection and, if so, what underlying mechanism is possible. METHODS: Caco-2 cell monolayers were rapidly infected with EPEC strain E2348/69 and immediately mounted in Ussing chambers. The monolayers were exposed to LBP before, after, and simultaneously with EPEC infection. Short circuit current (Isc) was measured in the Ussing chamber. RESULTS: EPEC infection caused an increase in short circuit current that was reduced by preincubation with LBP ( < 0.01). No direct bactericidal effect was observed, but LBP reduced the attachment of EPEC to Caco-2 cells. CONCLUSION: LBP can play an important role in reducing the secretory change in response to EPEC infection, possibly through inhibition of its binding. However, the presence of the probiotic agent before the infection is necessary. In this setting, its role is more preventive rather than therapeutic.     

Interleukin 8 in the tracheobronchial aspirate of infants acts as a neutrophil chemotactic factor in the development of chronic lung disease

BACKGROUND: We have already reported that there are some periods when a high interleukin 8 (IL-8) concentration is observed in the tracheobronchial aspirate of infants with chronic lung disease (CLD), although the changing pattern of the IL-8 concentration varies depending on the type of CLD. Interleukin 8 is known as a neutrophil chemotactic agent. Therefore, we asked whether IL-8 is an important neutrophil chemotactic factor in the tracheobronchial aspirate of infants who later develop CLD. METHODS AND RESULTS: We measured the neutrophil chemotactic activity of the tracheobronchial aspirate in CLD infants with or without anti-IL-8 antibody. Preincubation with anti-IL-8 immunoglobulinG resulted in a significant reduction of neutrophil chemotactic activity in the tracheobronchial aspirate. In infants with CLD following respiratory distress syndrome, there was a significant relationship between the IL-8 concentration and the neutrophil chemotactic activity of tracheobronchial aspirate without anti-IL-8 antibody, although no significant relationship was seen in infants with CLD following intra-uterine infection or with other CLD. CONCLUSIONS: Interleukin 8 in the tracheobronchial aspirate seems to play a significant role in recruiting neutrophils into the airways of patients with CLD, especially CLD following respiratory distress syndrome. We believe that in this type of CLD, IL-8 in the lung is generated as a result of hyperoxia rather than infection. In this situation, production of other neutrophil chemotactic factors or some factors that inhibit IL-8 activity may be insignificant.     

Eosinophils and neutrophils of human neonates have similar impairments of quantitative up-regulation of Mac-1 (CD11b/CD18) expression in vitro

Neonatal neutrophils have Mac-1-related functional abnormalities that may contribute to the enhanced susceptibility of neonates to bacterial infections. We developed a flow cytometric method using differences in light scattering and CD16 expression to distinguish eosinophils from neutrophils and studied their responses to stimulation in vitro. Mean forward light scatter was 20-60% greater for adult neutrophils than eosinophils, p less than or equal to 0.001, and side scatter was 30-60% greater for eosinophils than neutrophils, p less than or equal to 0.01, both before and after stimulation. Light scatter of neonatal and adult eosinophils did not differ. Mac-1 expression on adult eosinophils increased 60% after warming without specific stimulation and further increased 5-fold after incubation with 1 microM A23187, p less than 0.0001. Platelet activating factor and recombinant C5a produced modest increases in eosinophil Mac-1 expression, while N-formyl-met-leu-phe and leukotriene B4 had minimal effects. Histamine and the eosinophil chemotactic factor of anaphylaxis tetrapeptides had no effect on either Mac-1 or CR1 (CD35) expression. After stimulation with 1 microM A23187, eosinophils from eight healthy neonates at term expressed less Mac-1 than eosinophils from eight adults (mean +/- SD: 60.0 +/- 17.2 versus 89.1 +/- 12.8, p = 0.001) by the same ratio as neonatal to adult neutrophils (121.5 +/- 17.9 versus 178.2 +/- 11.7, p = 0.0004). We conclude that neonatal eosinophils and neutrophils have similar impairments of quantitative Mac-1 up-regulation. Further studies are needed to determine if neonatal eosinophils also have functional abnormalities related to Mac-1.     

Expression of the complement receptors CR1 and CR3 and the type III Fc gamma receptor on neutrophils from newborn infants and from fetuses with Rh disease

Developmental defects in neutrophil function, including diminished expression of plasma membrane receptors, may play an important role in the susceptibility of the newborn infant to infection. We used monoclonal antibodies and flow cytometry to study the expression of complement receptor type one (CR1), complement receptor type three (CR3), and Fc gamma receptor type three (FcRIII) on neutrophils from six fetuses with Rh disease, 10 preterm infants, nine term infants, and nine adults. Expression of the complement receptors on unstimulated cells was similar for all groups, but significant differences in complement receptor expression were observed after stimulation with N-formyl-methionyl-leucyl-phenylalanine (FMLP). Fetal, preterm, and term infant neutrophils expressed less CR3 than FMLP-stimulated neutrophils of adults [61 +/- 2, 48 +/- 4, and 66 +/- 4% (mean +/- SEM) of the mean for adults, p less than 0.05]. FMLP-stimulated CR1 expression for these groups was 61 +/- 6, 73 +/- 6, and 91 +/- 9% of the adult mean (p less than 0.05, fetal versus term infant and adult). Expression of both CR3 and CR1 increased with postconceptional age in the infants (r2 = 0.49, p less than 0.001 for CR3; r2 = 0.23, p less than 0.05 for CR1). Neutrophils of the preterm and term infants expressed less FcRIII than adult neutrophils (68 +/- 10 and 77 +/- 7% of the adult mean, p less than 0.05, for FMLP-stimulated cells), whereas fetal neutrophil FcRIII expression did not differ from that of the adult.(ABSTRACT TRUNCATED AT 250 WORDS)     

Expression and regulation of L-selectin on eosinophils from human adults and neonates.

L-Selectin, previously known as LEC.CAM-1, LECAM-1, LAM1, and as the MEL-14, Leu-8, TQ1, and DREG-56 antigens, is a leukocyte membrane protein that participates in adhesion to endothelium. We studied its expression on eosinophils using flow cytometry and the MAb Dreg-56 and Leu-8. Unstimulated peripheral blood eosinophils from healthy adults expressed about one third the level of L-selectin as neutrophils (mean +/- SD specific fluorescence: 20.9 +/- 3.2 versus 54.5 +/- 8.4, p = 0.0001, n = 18). After stimulation with A23187, L-selectin expression on eosinophils was rapidly lost. This was temporally correlated with increased expression of Mac-1 (CD11b/CD18); the kinetics on eosinophils and neutrophils were similar. Eosinophil expression of L-selectin decreased modestly after stimulation with platelet activating factor, but was minimally affected by N-formyl-methionyl-leucyl-phenylalanine, leukotriene B4, or C5a compared with their effects on neutrophils. Eosinophils from cord blood of healthy neonates born at term expressed less L-selectin than adult eosinophils (10.4 +/- 3.8 versus 19.4 +/- 2.7, p = 0.0001, n = 9); the relative reduction was the same as on cord blood neutrophils (36.4 +/- 8.2 versus 55.5 +/- 4.8, p = 0.0001, n = 9). Relative to baseline expression, the responses of neonatal and adult cells to stimulation did not differ. We conclude that neonatal eosinophils have abnormalities in L-selectin expression similar to neonatal neutrophils and suggest that decreased expression of L-selectin and a diminished responsiveness to direct stimulation with chemotactic factors are possible mechanisms that may limit the exudation of eosinophils.