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Department of Microbiology and Immunobiology Vaccine Center, University of Alabama, Birmingham, AL 35294;
Schering-Plough Research Institute, Kenilworth, NJ 07033; and
DNAX Research Institute of Cellular and Molecular Biology, Palo Alto, CA 93404
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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TCR-positive (TCR+) intraepithelial lymphocytes. Lptn is
chemotactic for NK and T cells and likely plays an important role in
maintaining the integrity of the epithelium and in mucosal immune
responses. In this study, we characterized the immune responses to OVA
given intranasally with Lptn to mice. This regimen enhanced
OVA-specific serum Ab responses and Ab titers in mucosal secretions.
Lptn also enhanced OVA-specific Ab-forming cells in mucosal and
systemic compartments. CD4-positive (CD4+) T cells isolated
from mucosal compartments and spleens of mice intranasally immunized
with OVA plus Lptn displayed higher OVA-specific proliferative
responses and greater synthesis of IFN-, IL-2, IL-4, IL-5, IL-6, and
IL-10 than did CD4+ T cells from mice given OVA without
Lptn. These studies indicate that Lptn has adjuvant properties and
suggest that Lptn present in the mucosa has the potential to enhance
mucosal and systemic Ab responses through help provided by Th1- and
Th2-type cells to link the initial innate signals of the mucosa with
the acquired immune system. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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TCR positive (TCR+) IEL is
centered around and in contact with six epithelial cells 2 . IELs
possess cytotoxic functions for protection against intracellular
infections and regulate humoral immune responses 3 . Recent evidence
suggests that IEL T cells provide signals (e.g., cytokines and
chemokines) to regulate and stabilize surrounding immune cells 4 .
Chemokines are chemotactic proteins produced by a variety of lymphoid
as well as epithelial cell types and are characterized by cysteine (C)
residues that are important for their structure and function.
Chemokines are often chemotactic and mitogenic for specific immune
cells. Of the four classes of chemokines, e.g., C, CC, CxC, and Cx3C,
lymphotactin (Lptn) is of the C type and is found in both mice and
humans 5, 6 . Lptn is similar to CC chemokines but differs by lacking
the first and third C residues, which are hallmarks of CC and CxC
chemokines 7 . Lptn produced by TCR+ IELs is
chemotactic for T and NK cells, but not for monocytes, neutrophils, or
dendritic cells 5, 8, 9, 10 . Moreover, memory T cells have been shown to
migrate in response to Lptn across HUVECs 11 . These properties,
together with the fact that Lptn is the most abundant chemokine
produced by a V3V1 TCR+ IEL
cell line 8 , suggest that this C chemokine could play a significant
role in mucosal immunity, inflammation, and/or tolerance.
While Lptn is known to enhance tumor immunity, presumably through the chemotaxis of NK and T cells 12 , its effects on mucosal immunity have not yet been evaluated. In this study, we investigated the hypothesis that Lptn may provide signals to link the initial innate system of the mucosa with the acquired mucosal immune system. To determine the immunological contribution that Lptn makes toward mucosal and systemic immunity, we investigated the effect of Lptn on Ag-specific immune responses.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Murine Lptn was produced by recombinant techniques and purified as previously described 13 . The identity and purity of the elutriated protein were confirmed by high pressure liquid chromatography, mass spectrophotometry, and amino acid sequence analysis. The endotoxin level quantified by the chromogenic Limulus amebocyte lysate assay (Cape Cod, Falmouth, MS) was <5 EU/mg. Therefore, subsequent doses (i.e., 0.01–5 µg) of Lptn given to mice resulted in endotoxin levels that were comparable to a dose of PBS, pH 7.5, alone. Chicken egg OVA and hen egg lysozyme were purchased from Sigma (St. Louis, MO).
Mice and immunizations
Female, 5- to 6-wk-old, C57BL/6 mice were procured from Charles River Laboratories (Wilmington, DE) and housed in horizontal laminar flow cabinets free of microbial pathogens. All mice used in this study were 8–12 wk of age. Following anesthesia, mice were intranasally immunized on days 0, 7, and 14 with 75 µg of OVA alone or with 0.01–5.0 µg of Lptn in 10 µl of PBS. In other studies, we have found that 75 µg of OVA is an optimal dose when coadministered with the mucosal adjuvant cholera toxin 14 . Experimental groups consisted of five mice, and studies were repeated three to five times. Control mice received 75 µg of hen egg lysozyme or PBS alone. The guidelines proposed by the committee for the Care of Laboratory Animal Resources Commission of Life Sciences, National Research Council were followed to minimize distress and pain of the mice.
Sample collection
Nasal and vaginal secretion samples were collected 1 wk following the third immunization (day 21) by washing the nasal or vaginal cavities three times with 50 µl (150 µl total) of PBS 14 . Saliva was obtained following i.p. injection with 100 µl of 500 ng/ml pilocarpine (Sigma) to induce saliva flow. Fecal samples were collected, weighed, and dissolved in PBS containing 0.1% sodium azide (e.g., 1 ml per 100 mg of fecal pellet). Following suspension by vortexing for 10 min, fecal samples were centrifuged, and supernatants were collected for analysis. Blood samples were collected by tail vein bleeding, and serum was obtained following centrifugation. Serum and mucosal secretions were collected on days 0, 7, 14, and 21 for OVA-specific Ab analysis by ELISA.
Ab isotype and IgG subclass analysis
The levels of OVA-specific Abs in saliva, fecal, nasal, vaginal,
and serum samples were measured by ELISA as previously described 14, 15 . Briefly, horseradish peroxidase (HRP)-conjugated-goat
anti-mouse 
, , or µ heavy chain-specific polyclonal Abs
adsorbed with human serum (Southern Biotechnology Associates,
Birmingham, AL) in 1% BSA in PBS containing 0.05% Tween 20 (B-PBS-T)
were used to detect murine Ab isotypes. Similarly, 100 µl of
biotin-conjugated rat anti-mouse 1 (G1–7.3 at 12.5 ng/ml),
2a (R19–15 at 125 ng/ml), 2b (R12–3 at 12.5 ng/ml), 3
(R40–82 at 50 ng/ml) and 
(G1–7.3; 1.25 µg/ml) (PharMingen, San
Diego, CA) heavy chain-specific mAbs were used to determine IgG
subclass and IgE isotype titers 15 . After incubation and washing, 100
µl of 0.5 µg/ml HRP-conjugated-anti-biotin mAb (Vector
Laboratories, Burlingame, CA) in B-PBS-T or 500 ng/ml polyHRP80
streptavidin (Research Diagnostics, Flanders, NJ) in PolyHRP Diluent
(Research Diagnostics) were added to IgG subclass or IgE detection
wells, respectively, and incubated for 3 h at room temperature.
Following incubation, the plates were washed six times, and the color
reaction was developed by adding 100 µl of 1.1 mM 2,2'-azino-bis
3 -ethylbenzthiazoline-6-sulfonic acid (Sigma) in 0.1 M
citrate-phosphate buffer, pH 4.2, containing 0.01%
H2O2 (ABTS solution). Endpoint titers were
expressed as the reciprocal log2 of the highest dilution,
which gave an optical density at 415 nm (OD415) of 
0.1 OD
unit above the OD415 of negative controls after a 20-min
incubation 16 .
Tissue disassociation and lymphoid cell isolation
Mice were sacrificed by CO2 inhalation 1 wk after the last immunization to quantify the OVA-specific Ab-forming cells (AFCs) and T cell responses present in immune compartments. Single-cell suspensions of spleen, mesenteric lymph nodes, Peyer’s patches, cervical lymph nodes, and salivary glands were prepared by aseptically removing tissues and then passing them through a sterile wire screen. The nasal-associated lymphoreticular tissue present in the nasal passage of mice was removed by scraping, and single-cell suspensions were obtained by passage over sterile glass wool 16 . Salivary gland, lower respiratory tract, and Peyer’s patch tissues were excised, minced, and further disrupted by incubation at 37°C with stirring in 0.3 mg/ml of collagenase type IV (Sigma) in RPMI 1640 (collagenase solution), as previously described 14 . Lamina propria tissues were removed, incubated at 37°C with stirring in 0.1 mM EDTA in PBS to separate IELs from lamina propria lymphocytes, and incubated in collagenase solution at 37°C with stirring to isolate lamina propria lymphocytes. Mononuclear cells in the collagenase solution were isolated at the 40–75% interface of a discontinuous Percoll gradient (Pharmacia, Uppsala, Sweden) 15 . Lymphocytes used ex vivo were maintained in complete medium, which consisted of RPMI 1640 supplemented with 10 ml/L of nonessential amino acids (Mediatech, Washington, DC), 1 mM sodium pyruvate (Sigma), 10 mM HEPES (Mediatech), 100 U/ml penicillin, 100 µg/ml streptomycin, 40 µg/ml gentamicin (Elkins-Sinn, Cherry Hill, NJ), 50 µM 2-ME (Sigma), and 10% FCS (Atlanta Biologicals, Norcross, GA).
Enzyme-linked immunospot (ELISPOT) assay
An ELISPOT assay was employed to detect total or OVA-specific
AFCs 16 . In brief, 96-well Millititer HA nitrocellulose-based plates
(Millipore, Bedford, MA) were coated with 100 µl of 1 mg/ml of OVA in
PBS, PBS only (negative control), or 0.5 µg/ml goat anti-mouse Ig
(H+L) human-adsorbed polyclonal Ab (Southern Biotechnology Associates)
and incubated overnight (12 h) at 4°C. Wells were subsequently
blocked with B-PBS for 2 h and washed. Whole cells were added to
wells in duplicate at 106, 5 x 105, and
105 cells/ml concentrations in complete medium and
incubated for 6 h at 37°C in 5% CO2. After washing
with PBS-T, individual AFCs were detected with
HRP-labeled-goat-anti-mouse -, µ-, or -chain specific Abs
(1 µg/ml; Southern Biotechnology Associates), visualized by adding
3-amino-9-ethylcarbazole buffer (Moss, Pasadena, MD) and counted using
a dissecting microscope (SZH Zoom Stereo Microscope System; Olympus,
Lake Success, NY).
Purification of CD4+ T cells and OVA-specific responses
T cells from the spleen, cervical lymph nodes, lower respiratory
tract, Peyer’s patches, and mesenteric lymph node tissues were
fractionated by passing single-cell suspensions over nylon wool for
1 h at 37°C. CD4+ T cells were enriched (>98%
purity) using Mouse CD4 Cellect Plus columns according to
manufacturer’s protocols (Biotex Laboratories, Edmonton, Alberta,
Canada). Next, purified CD4+ T cells were cultured at a
density of 5 x 106 cells/ml with 1 x
106 cells/ml of T cell-depleted and irradiated (3000 rads)
splenic feeder cells in complete medium containing OVA at 37°C in 5%
CO2. Purified T cells from nonimmunized mice were
stimulated with anti-CD3 mAb (145-2C11; PharMingen) or OVA as
positive or negative controls, respectively. To ascertain Ag-specific
proliferative responses, purified CD4+ T cells were
cultured in 96-well round-bottom plates. Following incubation for 3
days (as described above), cells were pulsed with 0.5 µCi of
methyl-3H-thymidine (Amersham, Arlington Heights, IL) per
well for 18 h. Cells were harvested on glass microfiber filter
paper (Whatman, Clifton, NJ), and radioactivity was determined by
liquid scintillation counting.
CD4+ T cell-derived cytokine analysis
For the assessment of cytokine production, 2 ml of culture
supernatants from 12-well flat-bottom plates were harvested after 5
days of incubation 14 . Control wells consisted of cells only or cells
cultured with BSA or 1 µg/ml Con A (Sigma). Cytokines in culture
supernatants were determined by ELISA as described previously 15 .
Briefly, Falcon 3912 Microtest plates were coated with 100 µl of 2.5
µg/ml of rat-anti-mouse IFN-, IL-2, IL-4, IL-5, IL-6, or IL-10
mAb (PharMingen) in 0.1 M bicarbonate buffer, pH 8.2, overnight at
4°C and blocked with 3% BSA in PBS. Next, 100 µl of serially
diluted recombinant murine cytokines as standards (PharMingen) or
culture supernatant samples were added in duplicate and incubated O/N
at 4°C. The plates were washed and then incubated with 0.2 µg/ml of
biotinylated secondary murine cytokine detection mAbs (PharMingen) in
B-PBS-T for 3 h at room temperature. After washing, wells were
incubated for 2 h in 100 µl of 0.5 µg/ml peroxidase-conjugated
anti-biotin mAb (Vector Laboratories) and developed with ABTS
solution, as described above. The cytokine ELISA was able to detect 15
pg/ml of IFN-, 5 pg/ml of IL-2, IL-4, and IL-5, 10 pg/ml of IL-6,
and 20 pg/ml of IL-10.
Statistics
The data are expressed as the mean ± SEM and compared using a two-tailed Student’s t test or an unpaired Mann Whitney U test. The results were analyzed using the Statview II statistical program (Abacus Concepts, Berkeley, CA) for Macintosh computers and were considered to be statistically significant if p values were <0.05. For cytokine levels of samples below the detection limit, levels were recorded as one-half the lower detection limit (e.g., 5 pg/ml for IL-6) for statistical analysis.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Our studies sought to determine whether the mucosal chemokine Lptn
possessed immunostimulatory activity for host responses to mucosal
immunization. We purposely used a relatively weak immunogen (i.e., OVA)
which when given intranasally to mice results in low Ab responses
without a mucosal adjuvant 14 . Increasing concentrations of Lptn were
then nasally administered as adjuvant with OVA. Administration of OVA
alone elicited low Ag-specific serum IgM and IgG Ab responses (Fig. 1
); however, 7 days following the second
immunization, mice intranasally immunized with OVA-plus-Lptn displayed
significantly higher (p < 0.05) serum titers
of anti-OVA IgA, IgM, and IgG Abs (Fig. 1). Groups receiving 0.1
µg of Lptn also displayed increases in OVA-specific serum IgE titers.
The increase in serum anti-OVA IgA and IgE Ab titers observed on
day 14 remained essentially the same through day 21 (Fig. 1). By day
21, Ag-specific IgM in all groups receiving OVA increased but the
highest serum Ab responses to OVA were of the IgG isotype (Fig. 1).
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). By the third dose of this
immunization schedule, serum anti-OVA IgG1 > IgG2b >
IgG2a > IgG3 Ab responses were at their highest levels (Fig. 2).
While the optimal serum OVA-specific IgG2a Ab titers occurred at day
21, the analogous IgG3 Ab titers peaked at day 14 and were sustained
through day 21 (Fig. 2). These results show that intranasal
administration of Lptn enhanced OVA-specific serum Ab responses in a
dose-dependent manner.
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Because Lptn enhanced serum Ab responses when coadministered
intranasally with Ag, we next assessed OVA-specific Ab titers in
mucosal secretions. Two intranasal doses of >1.0 µg of Lptn given
with OVA led to increases in Ag-specific IgA Ab titers in nasal wash
samples and fecal extracts at day 14. However, a third nasal
immunization with concentrations of Lptn 1 µg induced modest
OVA-specific IgG Abs in nasal and vaginal washes as well as saliva and
fecal samples (Fig. 3). Secretory IgA
(S-IgA) Ab levels in fecal samples on day 14 were among the highest
evaluated (data not shown) and remained high through day 21 (Fig. 3).
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). Increases in Ag-specific IgM
AFCs were seen in splenocytes, while lower respiratory tract-derived
cells displayed low to moderate increases in IgM and IgG anti-OVA
AFCs (Fig. 4). The number of OVA-specific IgG and IgM AFCs from these
tissues accounted for 20–40% of the total number of IgG and IgM AFCs.
The levels of Ag-specific IgG Ab titers in fecal extracts did not
correlate with ELISPOT analysis of cells isolated from intestinal
lamina propria tissues. Taken together, these findings indicate that
nasal delivery of Lptn enhanced Ag-specific serum IgA, IgM, and IgG
(i.e., IgG1 > IgG2b > IgG2a > IgG3) Abs and
up-regulated anti-OVA IgA and IgG Abs in mucosal secretions.
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Because Lptn induced systemic and mucosal Ab responses, we asked
whether this C chemokine promoted differential Ag-specific Th cytokine
profiles and T cell proliferative responses. We examined the
OVA-specific responses of CD4+ T cells isolated from the
lower respiratory tract, cervical lymph nodes, Peyer’s patches,
mesenteric lymph nodes, and spleen tissues. Because both low (0.01–0.1
µg) and high (1.0–5.0 µg) doses of Lptn with OVA enhanced the
Ag-specific cytokine and proliferative responses of T cells similarly,
mice receiving 1.0 µg of Lptn with OVA were further evaluated to
characterize Ag-specific cytokine profiles and proliferative
responses induced by Lptn. The CD4+ T cells isolated from
these immune compartments showed marked increases in OVA-specific
proliferative responses (Fig. 5).
Mesenteric lymph node- and spleen-derived CD4+ T cells from
mice that received Lptn showed the highest increases in
anti-OVA-mediated T cell proliferation.
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, IL-2, IL-4, IL-5, IL-6, and IL-10 in cultures (Fig. 5
). The Th
cell subpopulations from mice that did not receive Lptn showed cytokine
levels that were either low or below levels of detection. Ag-specific
CD4+ T cells of the mice that received Lptn with OVA
displayed considerably higher OVA-specific proliferative responses and
Th1- and Th2-type cytokine induction than did control groups (Fig. 5). |
Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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The role of Lptn in humoral immunity and the generation of Th cell
subset responses had been incompletely understood and unresolved until
now. Lptn coadministered with OVA induced brisk mucosal Ag-specific
S-IgA and serum anti-OVA Ab responses as well as increases in
OVA-specific serum IgG subclass Ab responses in a dose-dependent
manner. Additional experiments are needed to determine whether the
exogenous amounts of Lptn used in our model correlate with the in vivo
expression of Lptn in response to infection. However, it is important
to mention that even low doses of Lptn (0.01 µg) were sufficient to
up-regulate anti-OVA S-IgA Ab levels in mucosal secretions (Fig. 3)
as well as augment OVA-specific systemic Ab responses (Fig. 1) and
increase CD4+ T cell proliferative responses and cytokine
secretion (data not shown). The enhanced IgG subclass Ab responses can
be attributed to our use of a soluble protein Ag 17 as well as a
mixed Th2- and Th1-type of cytokine help 18, 19, 20 provided by
CD4+ T cells from mucosal and systemic immune compartments,
as noted by serum OVA-specific IgG1 followed by IgG2b, IgG2a,
and IgG3 Ab titers. Additionally, the anti-OVA IgE Abs
characterized were consistent with the Ag-specific induction of IL-4
cytokine levels produced by CD4+ T cells 19, 21, 22 . IL-4
promotes isotype switching to both IgG1 and IgE 23 and is required
for the maintenance of IgE 24 . Perhaps the IL-5 we observed from
Ag-stimulated CD4+ T cells helped to enhance the Ig Ab
levels measured in serum and mucosal secretion samples; in fact, this
cytokine has been shown to increase Ig secretion of IgA, IgG1, and IgE
committed B cells 25, 26 . IFN- production would support Th1-type
cytokine help for IgG2a and IgG3 Ab responses 18, 27, 28 . Low doses
of IFN- (e.g., 1500 units) have been shown to increase IgG2a
production in vivo, while considerably higher doses (e.g., 12,500
units) were required to induce decreases in IgG1 and IgE responses 29, 30 . Further, IFN- inhibits polyclonal IgG1 Ab responses rather than
Ag-specific IgG1 responses 30, 31 , which may partially explain why we
observed relatively high OVA-specific IgG1 Ab titers despite the
presence of IFN--secreting CD4+ T cells. While the
effects of IL-10 on murine B cell isotype switching are limited,
this Th2-type cytokine can inhibit Th1-type cytokine secretion patterns
in high concentrations 32 and has been shown to enhance Ig Ab
secretion of IgM-, IgG-, and IgA-committed human tonsillar B cells
23 . Clearly, both serum and mucosal Ag-specific Ab responses were
enhanced as a result of the effects of Lptn. Thus, the milieu of Th1-
and Th2-type cytokines produced by OVA-specific CD4+ T
cells from Lptn- plus OVA-immunized mice supports the Ig isotype and
IgG subclass Ab profiles observed in serum. Perhaps the
CD4+ T cell-derived Th2-type cytokines secreted by
OVA-cultured T cells from mucosal compartments supported the mucosal
S-IgA Ab responses observed.
The precise cytokine signals required for the induction of S-IgA Abs are not completely understood; however, previous studies have shown that the maintenance of mucosal IgA responses require Th2 cell-derived cytokines (e.g., IL-4, IL-5, IL-6, and IL-10) 15, 33, 34, 35 . The levels of the particular cytokine(s) required for B cells to express the IgA isotype were obviously provided in our experimental model. While IL-4, IL-5, and IL-6 do not induce IgA switching 36, 37, 38 , IL-5 and IL-6 induce surface IgA+ B cells to secrete IgA 33 . The cytokines produced by CD4+ T cells in mucosal and systemic compartments after local administration of Lptn partially explain why statistically significant (p < 0.05) increases in OVA-specific S-IgA occurred in mucosal secretions. ELISPOT analysis is particularly important for determining the source of anti-OVA IgG Abs in lymphoid tissues associated with the nasal tract and the salivary glands and nasal-associated lymphoreticular tissue (i.e., cervical lymph nodes). Additionally, there is no known active transport mechanism for IgG secretion by epithelial cells into the lumen of the gut; thus, the increase in anti-OVA IgG Ab titers in vaginal, nasal, and fecal samples may be the result of either salivary IgG or an influx of IgG from the periphery. Serum IgA contains both monomeric and polymeric IgA (pIgA), while mucosal IgA is pIgA. The production of S-IgA requires necessary signals to induce pIgA synthesis in the lamina propria and to efficiently transport S-IgA through the epithelium. The differences in serum IgA compared with mucosal IgA levels may be best explained by the characteristics of mucosal vs systemic IgA Ab responses. While Lptn provided the necessary signals for B cells to switch to IgA+ and IgA AFCs, this chemokine may not have provided the appropriate signals for optimal transport of S-IgA Abs from the intestinal lamina propria.
Our studies clearly show that Lptn enhances Ag-specific immune
responses and suggest that this C chemokine plays an important role in
mucosal immunity. The mechanisms that adjuvants employ to boost immune
responses are only partially understood. The most widely studied
adjuvants that enhance mucosal immunity when given nasally or orally
are cholera toxin, Escherichia coli heat labile toxin, and
related molecules (i.e., classical mucosal adjuvants). Cholera toxin
and related molecules have been shown to enhance Ag presentation by
APCs (i.e., monocytes, B cells, and epithelial cells) 39, 40 . Our
laboratory and others have demonstrated that classical mucosal
adjuvants may facilitate their adjuvant functions by up-regulating B7
and B7 ligand expression on APCs and T cells, respectively 40, 41, 42 .
Mucosal adjuvants also enhance immune responses by stimulating cytokine
release 34, 39 . However, the mechanisms that mucosal adjuvants use to
induce high responses in the gastrointestinal and female reproductive
tracts are not completely understood. It is possible that classical
mucosal adjuvants and Lptn affect homing characteristics of lymphoid
cells. Lptn has been shown to attract NK and T in vivo 13 , which
could partially explain its adjuvant effects. Presumably, the ability
of Lptn to attract NK and T cells could optimize Ag presentation and
therefore enhance the immunogenicity of coadministered Ag. On the other
hand, previous studies suggest that chemokine expression is under the
control of cytokines 43 ; however, recent studies have shown that
monocyte chemoattractant protein-1 can increase IL-4 production
while macrophage inhibitory protein-1 down-regulates this
cytokine in vitro 44 . Interestingly, both of these CC chemokines were
able to augment IFN- secretion of Con A-stimulated primary cultures
44 . Our data now provide in vivo support that chemokines can
influence the Th cytokine secretion patterns of CD4+ T
cells.
We have shown that nasal administration of Lptn can enhance acquired mucosal and systemic immunity. It remains for future studies to specify the precise contributions that Lptn makes toward the generation and differentiation of Ag-specific Ab and Th cell responses as well as immune cell interactions (i.e., costimulatory molecule regulation). However, the results presented here contribute to our understanding of the role Lptn plays in adaptive immunity. We have shown that Lptn can enhance the production of Ag-specific CD4+ T cells that produce Th1- and Th2-type cytokines in both systemic and mucosal immune compartments to support humoral immunity. Thus, Lptn acts as an innate mucosal adjuvant for the induction of adaptive immunity.
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Acknowledgments |
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Jerry R. McGhee, Department of Microbiology and Immunobiology Vaccine Center, University of Alabama, Room 273A BBRB, 845 19th Street South, Birmingham, AL 35294-2170. E-mail address:
3 Abbreviations used in this paper: IEL, intraepithelial lymphocyte; AFC, Ab-forming cells; ELISPOT, enzyme-linked immunospot; HRP, horseradish peroxidase; Lptn, lymphotactin; S-IgA, secretory-IgA.
Received for publication July 9, 1998. Accepted for publication November 2, 1998.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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K. Muller, S. Bischof, F. Sommer, M. Lohoff, W. Solbach, and T. Laskay Differential Production of Macrophage Inflammatory Protein 1{gamma} (MIP-1{gamma}), Lymphotactin, and MIP-2 by CD4+ Th Subsets Polarized In Vitro and In Vivo Infect. Immun., November 1, 2003; 71(11): 6178 - 6183. [Abstract] [Full Text] [PDF]
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L. Stievano, V. Tosello, N. Marcato, A. Rosato, A. Sebelin, L. Chieco-Bianchi, and A. Amadori CD8+{alpha}{beta}+ T Cells That Lack Surface CD5 Antigen Expression Are a Major Lymphotactin (XCL1) Source in Peripheral Blood Lymphocytes J. Immunol., November 1, 2003; 171(9): 4528 - 4538. [Abstract] [Full Text] [PDF]
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U. P. Singh, S. Singh, D. D. Taub, and J. W. Lillard Jr. Inhibition of IFN-{gamma}-Inducible Protein-10 Abrogates Colitis in IL-10-/- Mice J. Immunol., August 1, 2003; 171(3): 1401 - 1406. [Abstract] [Full Text] [PDF]
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P. N. Boyaka, A. Tafaro, R. Fischer, S. H. Leppla, K. Fujihashi, and J. R. McGhee Effective Mucosal Immunity to Anthrax: Neutralizing Antibodies and Th Cell Responses Following Nasal Immunization with Protective Antigen J. Immunol., June 1, 2003; 170(11): 5636 - 5643. [Abstract] [Full Text] [PDF]
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 J. W. Lillard Jr, U. P. Singh, P. N. Boyaka, S. Singh, D. D. Taub, and J. R. McGhee MIP-1alpha and MIP-1beta differentially mediate mucosal and systemic adaptive immunity Blood, February 1, 2003; 101(3): 807 - 814. [Abstract] [Full Text] [PDF]
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P. N. Boyaka, M. Ohmura, K. Fujihashi, T. Koga, M. Yamamoto, M.-N. Kweon, Y. Takeda, R. J. Jackson, H. Kiyono, Y. Yuki, et al. Chimeras of Labile Toxin One and Cholera Toxin Retain Mucosal Adjuvanticity and Direct Th Cell Subsets Via Their B Subunit J. Immunol., January 1, 2003; 170(1): 454 - 462. [Abstract] [Full Text] [PDF]
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 H. Huang, F. Li, J. R. Gordon, and J. Xiang Synergistic Enhancement of Antitumor Immunity with Adoptively Transferred Tumor-specific CD4+ and CD8+ T Cells and Intratumoral Lymphotactin Transgene Expression Cancer Res., April 1, 2002; 62(7): 2043 - 2051. [Abstract] [Full Text] [PDF]
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 U. F. Power, T. Huss, V. Michaud, H. Plotnicky-Gilquin, J.-Y. Bonnefoy, and T. N. Nguyen Differential Histopathology and Chemokine Gene Expression in Lung Tissues following Respiratory Syncytial Virus (RSV) Challenge of Formalin-Inactivated RSV- or BBG2Na-Immunized Mice J. Virol., December 15, 2001; 75(24): 12421 - 12430. [Abstract] [Full Text] [PDF]
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H. F. Staats, C. P. Bradney, W. M. Gwinn, S. S. Jackson, G. D. Sempowski, H.-X. Liao, N. L. Letvin, and B. F. Haynes Cytokine Requirements for Induction of Systemic and Mucosal CTL After Nasal Immunization J. Immunol., November 1, 2001; 167(9): 5386 - 5394. [Abstract] [Full Text] [PDF]
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