Absence of cross‐reactivity between murine Ly‐6C and Ly‐6G

Nagendra, Sanjai; Schlueter, Annette J.

doi:10.1002/cyto.a.20007

Cited by 48 publications

(41 citation statements)

References 29 publications

(25 reference statements)

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“…Thus, in addition to the depletion of neutrophils in blood and the injured spinal cord, it is possible that the anti-Ly6G/Gr-1 antibody might have depleted monocytoid cells and lymphocytes. Others, however, have not found the RB6-8C5 clone to cross-react with Ly6G (Nagendra and Schlueter, 2004). In our study, we were unable to document obvious changes to monocyte or lymphocyte numbers in blood after SCI as a result of antibody treatment (Fig.…”

Section: Discussioncontrasting

confidence: 78%

“…Ly6G is the major antigen on mature neutrophils identified by the RB6-8C5 clone (Hestdal et al, 1991;Fleming et al, 1993); the antibody depletes neutrophils in both blood and spleen for up to 2-3 d after injection (Conlan and North, 1994;Rakhmilevich, 1995). The RB6-8C5 anti-Ly6G/Gr-1 antibody, however, has been reported to bind to Ly6C, which is found on a subset of monocytoid cells and lymphocytes (Fleming et al, 1993;Daley et al, 2008), although this cross-reactivity was not observed by others (Nagendra and Schlueter, 2004).…”

Section: Methodsmentioning

confidence: 98%

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Depletion of Ly6G/Gr-1 Leukocytes after Spinal Cord Injury in Mice Alters Wound Healing and Worsens Neurological Outcome

Stirling¹,

Liu²,

Kubes³

et al. 2009

J. Neurosci.

200

155

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Spinal cord injury (SCI) induces a robust inflammatory response and the extravasation of leukocytes into the injured tissue. To further knowledge of the functions of neuroinflammation in SCI in mice, we depleted the early arriving neutrophils using an anti-Ly6G/Gr-1 antibody. Complete blood counts revealed that neutrophils increased ϳ3-fold over uninjured controls and peaked at 6 -12 h after injury, and that anti-Ly6G/Gr-1 treatment reduced circulating neutrophils by Ͼ90% at these time points. Intravital and spinning disk confocal microscopy of the exposed posterior vein and postcapillary venules showed a significant reduction in rolling and adhering neutrophils in vivo after anti-Ly6G/Gr-1 treatment; this was accompanied by a parallel reduction in neutrophil numbers within the injured spinal cord at 24 and 48 h as determined by flow cytometry. The evolution of astrocyte reactivity, a wound healing response, was reduced in anti-Ly6G/Gr-1-treated mice, which also had less spared white matter and axonal preservation compared with isotype controls. These histological outcomes may be caused by alterations of growth factors and chemokines important in promoting wound healing. Importantly, anti-Ly6G/Gr-1 treatment worsened behavioral outcome as determined using the Basso Mouse Scale and subscores. Although the spectrum of cells affected by anti-Ly6G/Gr-1 antibody treatment cannot be fully ascertained at this point, the correspondence of neutrophil depletion and worsened recovery suggests that neutrophils promote recovery after SCI through wound healing and protective events that limit lesion propagation.

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Section: Discussioncontrasting

confidence: 78%

Section: Methodsmentioning

confidence: 98%

Depletion of Ly6G/Gr-1 Leukocytes after Spinal Cord Injury in Mice Alters Wound Healing and Worsens Neurological Outcome

Stirling¹,

Liu²,

Kubes³

et al. 2009

J. Neurosci.

200

155

View full text Add to dashboard Cite

show abstract

“…This seems to support the idea that anti-Gr-1 mAb binds to Ly6C with minor avidity when compared with Ly6G, as originally suggested [11]. However, even if some groups have concluded that anti-Gr-1 mAb preferentially binds to either Ly6G or Ly6C, many discrepancies about epitope recognition and competition between Ly6C and Ly6G molecules have been reported [12,13], and Gr-1 expression has been largely used to distinguish monocyte subsets [14]. The F4/80 and CD68 markers, both associated with a monocyte/macrophage phenotype, were principally expressed by the Gr-1 int fraction.…”

Section: Mdsc Subset Phenotypesupporting

confidence: 80%

Hierarchy of immunosuppressive strength among myeloid‐derived suppressor cell subsets is determined by GM‐CSF

et al. 2009

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CD11b+/Gr‐1+ myeloid‐derived suppressor cells (MDSC) contribute to tumor immune evasion by restraining the activity of CD8+ T‐cells. Two major MDSC subsets were recently shown to play an equal role in MDSC‐induced immune dysfunctions: monocytic‐ and granulocytic‐like. We isolated three fractions of MDSC, i.e. CD11b+/Gr‐1high, CD11b+/Gr‐1int, and CD11b+/Gr‐1low populations that were characterized morphologically, phenotypically and functionally in different tumor models. In vitro assays showed that CD11b+/Gr‐1int cell subset, mainly comprising monocytes and myeloid precursors, was always capable to suppress CD8+ T‐cell activation, while CD11b+/Gr‐1high cells, mostly granulocytes, exerted appreciable suppression only in some tumor models and when present in high numbers. The CD11b+/Gr‐1int but not CD11b+/Gr‐1high cells were also immunosuppressive in vivo following adoptive transfer. CD11b+/Gr‐1low cells retained the immunosuppressive potential in most tumor models. Gene silencing experiments indicated that GM‐CSF was necessary to induce preferential expansion of both CD11b+/Gr‐1int and CD11b+/Gr‐1low subsets in the spleen of tumor‐bearing mice and mediate tumor‐induced tolerance whereas G‐CSF, which preferentially expanded CD11b+/Gr‐1high cells, did not create such immunosuppressive environment. GM‐CSF also acted on granulocyte–macrophage progenitors in the bone marrow inducing local expansion of CD11b+/Gr‐1low cells. These data unveil a hierarchy of immunoregulatory activity among MDSC subsets that is controlled by tumor‐released GM‐CSF.

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“…In this study, Ly-6G was used as a marker for granulocytic differentiation. An antigranulocyte receptor 1 Ab (RB6-8C5), which has been widely used for the detection of mouse granulocytes (21,25,26), recognizes both Ly-6G and Ly-6C (22,27). However, the expression of Ly-6C is not restricted to granulocytes, and so, the Ab crossreacts with nongranulocytic lineages such as dendritic cells (28), monocytes (29,30), and a subpopulation of lymphocytes (31).…”

Section: Discussionmentioning

confidence: 99%

C/EBPβ Is Involved in the Amplification of Early Granulocyte Precursors during Candidemia-Induced “Emergency” Granulopoiesis

Satake

Hirai

Hayashi

et al. 2012

The Journal of Immunology

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Granulopoiesis is tightly regulated to meet host demands during both “steady-state” and “emergency” situations, such as infections. The transcription factor CCAAT/enhancer binding protein β (C/EBPβ) plays critical roles in emergency granulopoiesis, but the precise developmental stages in which C/EBPβ is required are unknown. In this study, a novel flow cytometric method was developed that successfully dissected mouse bone marrow cells undergoing granulopoiesis into five distinct subpopulations (#1–5) according to their levels of c-Kit and Ly-6G expression. After the induction of candidemia, rapid mobilization of mature granulocytes and an increase in early granulocyte precursors accompanied by cell cycle acceleration was followed by a gradual increase in granulocytes originating from the immature populations. Upon infection, C/EBPβ was upregulated at the protein level in all the granulopoietic subpopulations. The rapid increase in immature subpopulations #1 and #2 observed in C/EBPβ knockout mice at 1 d postinfection was attenuated. Candidemia-induced cell cycle acceleration and proliferation of hematopoietic stem/progenitors were also impaired. Taken together, these data suggest that C/EBPβ is involved in the efficient amplification of early granulocyte precursors during candidemia-induced emergency granulopoiesis.

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Absence of cross‐reactivity between murine Ly‐6C and Ly‐6G

Cited by 48 publications

References 29 publications

Depletion of Ly6G/Gr-1 Leukocytes after Spinal Cord Injury in Mice Alters Wound Healing and Worsens Neurological Outcome

Depletion of Ly6G/Gr-1 Leukocytes after Spinal Cord Injury in Mice Alters Wound Healing and Worsens Neurological Outcome

Hierarchy of immunosuppressive strength among myeloid‐derived suppressor cell subsets is determined by GM‐CSF

C/EBPβ Is Involved in the Amplification of Early Granulocyte Precursors during Candidemia-Induced “Emergency” Granulopoiesis

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