Distinct locomotive patterns of granulocytes, monocytes and lymphocytes in a stable concentration gradient of chemokines

Abstract

The pattern of leukocyte locomotion can be changed in many pathological situations, but its accurate analysis is difficult because of technological limitation. In the present study, by using a newly developed time-lapse videomicroscopic technique, we have analyzed the locomotive patterns of leukocytes in a stable concentration gradient of chemokines. Granulocytes, monocytes, and lymphocytes were purified from adult human peripheral blood. Locomotive behavior of the leukocytes was analyzed by an optical assay using a microchannel producing a stable concentration gradient of chemokines. Videomicroscopic analysis showed distinct locomotive patterns of granulocytes, monocytes, and lymphocytes. Granulocytes were intrinsically motile, vigorously moving in random direction without any chemokine stimulation. Upon stimulation with CXCL8/IL-8, the speed of migration was increased from 13.3 +/- 2.8 to 19.4 +/- 2.5 microm/min (P = 0.002, n = 100) and they moved toward the chemokine, although many cells still wandered very much. Stimulation with CCL5/RANTES or CXCL12/SDF-1alpha induced similar changes in locomotive pattern. On the other hand, most lymphocytes did not polarize or move spontaneously without chemokine stimulation. Stimulation with CXCL12 induced directional migration in 37% of the lymphocytes at a speed of 9.6 +/- 1.6 microm/min (n = 100). The movement pattern of monocytes was similar to that of granulocytes in that they tend to become polarized and move spontaneously, but they moved at a very slow speed ranging from 3.9 to 4.2 microm/min even with chemokine stimulation. The new optical assay may be useful for many diagnostic as well as basic studies.

Migrating Myeloid Cells Sense Temporal Dynamics of Chemoattractant Concentrations

“…It is worth noting that many of the custom (Abhyankar et al, 2008; Amadi et al, 2010; Saadi et al, 2007) and commercially available gradient generators (EZ Taxiscan, Ibidi) (Bae et al, 2008; Kanegasaki et al, 2003) used in these experiments require cell introduction into the chamber, followed by gradient establishment. In these cases, there is a gradual increase in chemokine concentration throughout the device as the gradient develops until a steady state persistent stable gradient is established.…”

Misbalanced CXCL12 and CCL5 Chemotactic Signals in Vitiligo Onset and Progression

“…These lesions were also characterized by the accumulation of CXCL12 þ APC and the presence of ligand (CXCL12)-positive, receptor (CXCR4)-positive, and double-positive cells. Because CXCR4 is constitutively expressed in granulocytes, dendritic cells (DCs), and T cells (Bae et al, 2008;Beider et al, 2003;Stein and Nombela-Arrieta, 2005), it is plausible that melanocyte-derived CXCL12, along with other chemotactic signals, recruits these APCs, which in turn secrete more CXCL12, leading to accumulation of various leukocytes and polarization of the CXCR4 þ keratinocytes. Although the significance of KC polarization in the skin is not well understood, it is possible that CXCR4 signaling mitigates KC proliferation triggered by leukocyte-derived cytokines, because it has been shown in nonhyperplastic psoriatic skin regions (Takekoshi et al, 2013).…”

“…Studies of vitiligo in animal models showed that this chemokine plays an important role in the maintenance of CXCR3 þ T-cell effector function (Rashighi et al, 2014). Elevated levels of CXCL12 in epithelial tissues were reported in blister fluids after skin burns (Avniel et al, 2006), lichen planus (Ichimura et al, 2006), and cutaneous lupus erythematosus (Meller et al, 2005), where inflammatory and immune responses were supported by the recruitment of CXCR4-positive leukocytes (Bae et al, 2008;Beider et al, 2003). Autoimmune encephalomyelitis (Dos Santos et al, 2008) and thyroid disorders (Ferrer-Francesch et al, 2006) were linked to the elevated levels of CCL5.…”

In Vivo Imaging and Quantitative Analysis of Leukocyte Directional Migration and Polarization in Inflamed Tissue

“…We found that monocytes started their target-oriented interstitial migration later than neutrophils and migrated rather slower than neutrophils. Interestingly, monocytes have been reported to move slower than neutrophils along a stable chemotactic gradient in vitro [33] , [34] . It seems possible that the ability of neutrophils for fast movement would enable them to accumulate more rapidly at the site of inflammation.…”