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Review
. 1999 Feb;96(2):155-63.
doi: 10.1046/j.1365-2567.1999.00681.x.

Local immune responses in afferent and efferent lymph

D M Haig  1 , J Hopkins, H R Miller
Affiliations
Review

Local immune responses in afferent and efferent lymph

D M Haig et al. Immunology. 1999 Feb.
No abstract available

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Figures

Figure 1
Figure 1
Flow rates and cellular composition of lymph around single skin-draining lymph nodes of adult sheep. Afferent lymph contains antigen-presenting dendritic cells, T cells and a small proportion of B cells. Lymph flow and cell output rates are low. Efferent lymph contains >98% lymphocytes with a higher proportion of B cells and much higher flow and cell output rates than in afferent lymph.2–5 Greater than 90% of efferent lymphocytes are derived from the blood via high endothelial venules (HEV). L-selectin on lymphocytes is associated with adhesion to peripheral lymph node HEV. For the details of the cytokines studied to date, see Fig. 2 and the text.
Figure 2
Figure 2
Differential cytokine production in afferent and efferent lymph during an anamnestic immune response to 1 mg orf virus antigen given intra-dermally to previously infected sheep (refs 14, 21 and unpublished). Mean total and lymphoblast output for each group of five sheep is shown for comparison (top graphs). Lymph plasma contains activity generated in the skin (afferent lymph) or prefemoral lymph node (efferent lymph) as well as by cells in the lymph. Cytokine production by 4×106 lymph cells in 1 ml medium after culture for 18 hr in the absence of exogenous antigen was assayed in cell-free supernates (CFS) as a measure of the activation status of the cells following in vivo antigen challenge. Three examples of cytokines are shown. Note the difference in y-axis scales between afferent and efferent lymph. The chemokine IL-8 is produced within 12 hr of antigen challenge in both lymph compartments, most notably by afferent lymph cells. Low titres were detected in lymph plasma (there is no evidence for inhibitors in lymph plasma). GM-CSF is a haemopoietin active on macrophages, neutrophils and dendritic cells. Large quantities are produced within the lymph node and by afferent but not efferent lymph cells. This might indicate a consequence of accessory cell–lymphocyte interactions in afferent lymph and the lymph node. IFN-γ, like GM-CSF, is produced rapidly and at high titre by afferent lymph cells compared to efferent lymph cells where production is low and delayed by comparison. Production by cells correlated with the appearance of lymphoblasts. Differential and temporal appearance of the cytokines in the lymph compartments was similar in sheep reinfected with orf virus where CD4+/CD45RA− T lymphoblasts were the principal source of GM-CSF, IFN-γ (and IL-2).14,21 The results show a marked concentration of activated cytokine-producing cells in afferent lymph compared to efferent lymph during the anamnestic immune response, consistent with an activated effector/memory cell population in afferent lymph. A function for IL-8 (which precedes IL-1β detection14) in the initial phase of the anamnestic immune/inflammatory response is also implied.
Figure 3
Figure 3
Constitutive cytokine gene transcription by ‘resting’ afferent versus efferent CD4+ T cells. Lymph flow was allowed to settle for 10 days prior to harvesting the cells. 2×107 CD4+ T cells isolated from afferent or efferent lymph by magnetic activated cell sorting (MACS) (>94% purity) were incubated with 0·01mM of α-amatin to block mRNA translation. RNA from 1×107 CD4+ T cells was purified and subjected to reverse transcription–polymerase chain reaction (RT–PCR) using primers for APTase, ovine IL-4, IL-6, IL-10 and IFN-γ. Ten millilitre samples of amplified DNA were slot-blotted onto Hybond-N® using a Biodot® manifold and visualised by Southern hybridization with 32P-labelled cytokine-specific oligonucleotides (internal sequence). The results show that activated CD4+ T cells exhibiting type 1 (IFN-γ) and type 2 (IL-4, IL-6 and IL-10) cytokine expression are concentrated in afferent lymph.
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References

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