ライフサイエンスコーパス: microvesicle

1 rrestin domain-containing protein 1-mediated microvesicles.

2 matrix metalloprotease (MT1-MMP) to shedding microvesicles.

3 ells adopt an amoeboid phenotype and release microvesicles.

4 tes TCR sorting and release in extracellular microvesicles.

5 to generate T-cell antigen receptor-enriched microvesicles.

6 cancers, are often packaged within secreted microvesicles.

7 lar signals in response to isolated synaptic microvesicles.

8 Vif in lysosomes and by secretion of Vif in microvesicles.

9 the miRNA content of the macrophage-derived microvesicles.

10 te the binding of circulating leukocytes and microvesicles.

11 d from cells in association with proteins or microvesicles.

12 icrovesicles from nontumor host cell-derived microvesicles.

13 al similarities between viruses and cellular microvesicles.

14 immunogold-labeled intact and permeabilized microvesicles.

15 ere associated with both necrotic debris and microvesicles.

16 ain synaptic vesicles and PC12 synaptic-like microvesicles.

17 stases and analyzed tumor cells, stroma, and microvesicles.

18 et cells to which they are trafficked within microvesicles.

19 y resolved in animals cranially grafted with microvesicles.

20 triggering their secretion by extracellular microvesicles.

21 e desintegration of FlnA-null platelets into microvesicles, a process that occurs spontaneously durin

22 s support a scenario by which T cell-derived microvesicles act as intercellular carriers of functiona

23 ggest that the neuroprotective properties of microvesicles act through a trophic support mechanism th

24 Exosomes and related microvesicles also aid cells in exporting less-needed mo

25 Here, we show that tumour microvesicles also carry DNA, which reflects the genetic

26 h the release of a soluble growth factor and microvesicles, alter the type of particles engulfed by n

27 bound extracellular vesicles (EVs; exosomes, microvesicles and apoptotic bodies) containing proteins,

28 ys for protein delivery with both eukaryotic microvesicles and bacterial surface secretion systems.

29 ield (n = 30), respectively; (2) circulating microvesicles and blood cells; and (3) lungs from a mono

30 moted the biogenesis of onco-miR-221(hi) CAF microvesicles and established stromal CSC niches in expe

31 reviews the current knowledge on the role of microvesicles and exosomes from various cellular origins

32 most of the known cell-derived vesicles, the microvesicles and exosomes.

33 racellular vesicles (EVs): apoptotic bodies, microvesicles and exosomes.

34 losion of novel functions: the biogenesis of microvesicles and exosomes; plasma membrane wound repair

35 Here, we characterized macrophage-derived microvesicles and explored their role in the differentia

36 cells are able to switch between the use of microvesicles and invadopodia to facilitate invasion thr

37 ence of the kinin B1 receptor on endothelial microvesicles and its contribution to the inflammatory p

38 racellular vesicles (EVs), such as exosomes, microvesicles and large oncosomes, are involved in this

39 Furthermore, tumor microvesicles and miR-21 require c-Jun N-terminal kinase

40 and Alu retrotransposon elements, in tumour microvesicles and these transposable elements could be t

41 rovesicles from their cell surface (shedding microvesicles) and from internal, endosome-derived membr

42 cargo appears to be selectively sorted into microvesicles, and adhesion to the extracellular matrix

43 Extracellular vesicles (exosomes, microvesicles, and apoptotic bodies) are ubiquitous in h

44 rious extracellular vesicles (EVs; exosomes, microvesicles, and apoptotic bodies) that differ in biog

45 icles (EVs), which include apoptotic bodies, microvesicles, and exosomes, have emerged as important p

46 tracellular vesicles (EVs) such as exosomes, microvesicles, and large oncosomes are involved in tumor

47 ial effects by release of paracrine factors, microvesicles, and transfer of mitochondria, all of whic

48 Stem cell-derived microvesicles appear to be naturally equipped to mediate

49 ous report, the majority of peripheral blood microvesicles are derived from platelets, while mononucl

50 Thus, complement-coated RBC-derived microvesicles are elevated in HUS patients and induced i

51 Exosomes and microvesicles are extracellular nanovesicles released by

52 Tumor-derived microvesicles are heterogeneous membrane-bound sacs that

53 Microvesicles are membrane-bound vesicles released from

54 Microvesicles are small membrane-bound particles compris

55 omain-containing protein 1 (ARRDC1)-mediated microvesicles) are extracellular vesicles that bud direc

56 lular vesicles (EVs), including exosomes and microvesicles, are 30-800 nm vesicles that are released

57 ellular vesicles (EVs), namely, exosomes and microvesicles, are important mediators of intercellular

58 lular vesicles (EVs), including exosomes and microvesicles, are present in a variety of bodily fluids

59 which are derived from MVBs, ARRDC1-mediated microvesicles (ARMMs) lack known late endosomal markers.

60 Cancers have adapted the exosome and related microvesicles as a pathway by which neoplastic cells com

61 extracellular matrix (ECM) is facilitated by microvesicle-associated integrin receptors.

62 The functional roles of sCTLA-4 and microvesicle-associated Tm-CTLA-4 warrant further invest

63 es for the increased release of exosomes and microvesicles bearing M. tuberculosis peptide-MHC-II com

64 sed on our observations, we hypothesize that microvesicles bind to and activate target cells.

65 he vertical trafficking of cargo to sites of microvesicle biogenesis at the cell surface.

66 e describe Rho-mediated pathways involved in microvesicle biogenesis through the regulation of myosin

67 Despite growing understanding of microvesicle biogenesis, function and contents, mechanis

68 activation and the proteolytic activities of microvesicles, both of which are thought to correlate di

69 sferrin-positive endosomes and synaptic-like microvesicles but not in insulin-containing large dense

70 ophil chemotaxis, and the reduction of these microvesicles by C1-inhibitor should be explored as a po

71 We also show that circulating GBM microvesicles can be used to analyze primary tumor mutat

72 NA-suggests multipronged mechanisms by which microvesicles can condition the extracellular milieu to

73 rst demonstration that C. neoformans-derived microvesicles can facilitate cryptococcal traversal acro

74 these studies demonstrate the importance of microvesicle cargo sorting in matrix degradation and dis

75 s, the v-SNARE, VAMP3, regulates delivery of microvesicle cargo such as the membrane-type 1 matrix me

76 here, by membrane invagination, intraluminal microvesicles carrying membranal bioactive FasL and TRAI

77 s study, we found that C. neoformans-derived microvesicles (CnMVs) can enhance the traversal of the b

78 rum, Stx2 induced the release of RBC-derived microvesicles coated with C5b-9, a process that was inhi

79 release of B1 receptor-positive endothelial microvesicles compared with normal plasma, an effect sig

80 pCRP*-microvesicle complexes lead to enhanced recruitment of l

81 mean 72% decrease (P = 0.01) in C4d+/CD144+ microvesicle concentration compared with pretreatment va

82 We studied microvesicle concentration in the plasma of 95 kidney tr

83 by comparing interleukin 6 concentration and microvesicle concentration.

84 t also induced a shift in the size of plasma microvesicles, consistent with active release of microve

85 Thus, tumour microvesicles contain a repertoire of genetic informatio

86 Previously, we showed that plasma microvesicles contain microRNAs (miRNAs).

87 response to iron restriction and that these microvesicles contain mycobactin, which can serve as an

88 Microvesicles contain proteins and nucleic acids.

89 Tumour cells release an abundance of microvesicles containing a selected set of proteins and

90 Altogether, these data support a role for microvesicles contributing to T. cruzi evasion of innate

91 C4d+/AVB+ microvesicles correlated with AMR biopsy severity.

92 These microvesicles deliver transcellular signals across antig

93 It is unknown how exosomes/microvesicles deliver transmembrane proteins such as PMC

94 ivergent secretory organelles (synaptic-like microvesicles, dense-core vesicles, lysosomes, exosomes

95 nd functional studies reveal that IGF-1- and microvesicle-dependent communication between macrophages

96 MT1-MMP delivery to nascent microvesicles depends on the association of VAMP3 with t

97 Recently, we have shown that microvesicles derived from activated T cells (mvT*s) can

98 It has recently been shown that microvesicles derived from activated T cells can stimula

99 orting this hypothesis, by demonstrating how microvesicles derived from cancer-associated fibroblasts

100 iogenesis: one pool comprises 100- to 600-nm microvesicles derived from direct budding of the plasma

101 rent Plasmodium strains are known to produce microvesicles derived from the infected red blood cells

102 s from wild-type cells, B1 receptor-positive microvesicles derived from transfected human embryonic k

103 alytical technique for profiling circulating microvesicles directly from blood samples of patients wi

104 ic cells produce and release CrkI-containing microvesicles (distinct from exosomes and apoptotic bodi

105 igh levels of C3- and C9-bearing RBC-derived microvesicles during the acute phase, which decreased af

106 Endothelial microvesicles (EMVs) are elevated in patients with tradi

107 Animal cells bud exosomes and microvesicles (EMVs) from endosome and plasma membranes.

108 gger the release of infectious extracellular microvesicles (EMVs) that contain viral material.

109 Exosomes, also known as microvesicles (EMVs), are nano-sized membranous particle

110 me elements that target proteins to exosomes/microvesicles (EMVs), HIV, and other retrovirus particle

111 ll vesicles, otherwise known as exosomes and microvesicles (EMVs).

112 cesses, including formation of extracellular microvesicles, enveloped virus budding, and the abscissi

113 d, we found full-length IL-6R on circulating microvesicles, establishing microvesicle release as a no

114 membrane vesicles [OMVs], exosomes, shedding microvesicles, etc.), the conserved functions and mechan

115 Exosomes and other extracellular microvesicles (ExMVs) have important functions in interc

116 n adult retinal pigment epithelial cells via microvesicles (exosomes), independent of the endoplasmic

117 extracellular RNAs (exRNAs) associated with microvesicles, exosomes (collectively called EVs), and r

118 it with extracellular fractions enriched in microvesicles, exosomes and ribonucleoprotein complexes.

119 RECENT FINDINGS: Microvesicles, exosomes, apoptotic bodies, lipoproteins,

120 mata and the presence of TDP-43 oligomers in microvesicles/exosomes and show that microvesicular TDP-

121 MiR-126 was secreted in microvesicles/exosomes, and inhibition of their release

122 eview how exosomes and related extracellular microvesicles facilitate the progression and metastases

123 ovesicles, consistent with active release of microvesicles following liver injury.

124 Suppression of RhoA signaling blocks microvesicle formation but enhances the formation of inv

125 ne (an antidepressant agent known to inhibit microvesicle formation by interfering with membrane-asso

126 to each organism such as adherence proteins, microvesicle formation, toxin production and the propens

127 allooning was closely followed by a surge in microvesicle formation, which was absent when synchrony

128 Upon their release, microvesicles formed a complex on the T. cruzi surface w

129 were able to identify miR-155 in circulating microvesicles from both individuals with MBL and patient

130 Extracellular microvesicles from GBV-C E2-expressing cells contained E

131 ) and control plasma (n=6) on the release of microvesicles from glomerular endothelial cells.

132 Furthermore, microvesicles from mycobacterium-infected macrophages we

133 differentiate glioblastoma multiforme (GBM) microvesicles from nontumor host cell-derived microvesic

134 Importantly, microvesicles from patients with HTR metastatic disease

135 ls were significantly reduced in circulating microvesicles from patients with PAH and the lungs of th

136 RBCs and/or RBC-derived microvesicles from patients with STEC-HUS (n = 25) were

137 mechanism that results in direct budding of microvesicles from the plasma membrane, providing a form

138 rotein sorting 4 (VPS4) mediates scission of microvesicles from the T-cell plasma membrane.

139 Cells secrete various membrane-enclosed microvesicles from their cell surface (shedding microves

140 e of protease-loaded plasma membrane-derived microvesicles from tumor cells into the surrounding envi

141 We also find amplified c-Myc in serum microvesicles from tumour-bearing mice.

142 Compared with microvesicles from wild-type cells, B1 receptor-positive

143 In Syt IV knockouts, dense-core and microvesicle fusion was enhanced in cell-attached patche

144 sites, infected red blood cell (RBC)-derived microvesicles, gametocytes, and uninfected RBCs.

145 - and MAPK-dependent apoptosis and apoptotic microvesicle generation.

146 Microvesicles have a variety of cellular functions from

147 (EVs), which include exosomes and ectosomes/microvesicles, have emerged as important intercellular r

148 We determined that CAF-derived microvesicles horizontally transferred miR-221 to tumor

149 Exosomes and microvesicles (i.e., extracellular vesicles: EVs) have b

150 Thus, new and improved strategies for microvesicle identification, isolation, and capture will

151 dly higher levels of circulating endothelial microvesicles, identified by flow cytometry analysis, an

152 ic lymphocytic leukemia (CLL) B-cell-derived microvesicles in CLL plasma carry a constitutively phosp

153 Consequently, the results revealed a role of microvesicles in iron acquisition in M. tuberculosis, wh

154 sess the evidence for a role of exosomes and microvesicles in normal cardiovascular physiology, as we

155 fic and readily differentiates exosomes from microvesicles in samples containing 1000-fold excess of

156 d platelet-derived MPs are the most abundant microvesicles in the circulation.

157 les, an effect abrogated by reduction of the microvesicles in the perfused samples.

158 (ICAM1) is released from senescent cells by microvesicles independently of ADAM17.

159 patient samples, we show that tumor-derived microvesicles induce apoptosis of skeletal muscle cells.

160 Furthermore, we find that microvesicles induce the differentiation of macrophages.

161 Microvesicles induced the upregulation of several cluste

162 ong these, IL24 appeared to be a hallmark of microvesicle-induced activation.

163 Production of IL-24 is a unique feature of microvesicle-induced MC activation because its productio

164 The aim of this study was to characterize microvesicle-induced MC expression patterns.

165 osphocholine)-hexane), which blocks the pCRP-microvesicle interactions, abrogates these proinflammato

166 internalization of activated T cell-derived microvesicles into human MCs occurred within 24 hours.

167 portantly we show that TRAILshort is shed in microvesicles into the cellular microenvironment and the

168 ed large quantities of small, membrane-bound microvesicles into the circulation.

169 Microvesicles, introduced onto a dedicated microfluidic

170 A subgroup of such microvesicles is called exosomes and is described in blo

171 Finally, we describe functionally similar microvesicles isolated from bodily fluids of ovarian can

172 However, early studies of microvesicles (L-particles) secreted from herpes simplex

173 motaxis, an effect decreased by reduction of microvesicle levels and by blocking the B1 receptor.

174 ion with T. cruzi showed a rapid increase of microvesicle levels in mouse plasma, and infection with

175 metastases of cancers and describe how these microvesicles may affect clinical care.

176 Thus, B1 receptor-positive endothelial microvesicles may contribute to chronic inflammation by

177 For example, microvesicle-mediated genetic transfer can regulate the

178 Overall, our results illuminate how microvesicle-mediated horizontal transfer of genetic mat

179 nfected cells provided the first evidence of microvesicle-mediated intercellular communication.

180 Recent discoveries describing the microvesicle-mediated intercellular transfer of function

181 The hypothesis that microvesicle-mediated miRNA transfer converts noncancer

182 regulated release of the exosome but not the microvesicle MHC-II pool.

183 EVs (that comprise exosomes and microvesicles/microparticles) have a size ranging from 4

184 lular vesicles are classified into exosomes, microvesicles/microparticles, or apoptotic bodies, origi

185 cultures exhibit distinct compositions, with microvesicles most closely reflecting cellular transcrip

186 ound that acid exposure induced a remarkable microvesicle (MV) release from lung epithelium as detect

187 of patients with cancer contain membraneous microvesicles (MV) able to induce apoptosis of activated

188 These changes include rapid shedding of microvesicles (MV) and the nonconventional secretion of

189 Because microvesicles (MV) are biomarkers of endothelial dysfunc

190 Exosomes and microvesicles (MV) are cell membranous sacs originating

191 Microvesicles (MV) derived from human cancer cells have

192 varied widely in size, indicating that both microvesicles (MVs) and dense-core vesicles (DCVs) under

193 latelets, and tissue factor-positive (TF(+)) microvesicles (MVs) are all potential factors that alone

194 Microvesicles (MVs) are anuclear fragments of cells rele

195 described cerebrospinal fluid (CSF) myeloid microvesicles (MVs) as a marker of microglia activation

196 Accurate analysis of specific microvesicles (MVs) from biofluids is critical and chall

197 Microvesicles (MVs) have been indicated as important med

198 used this system to investigate the role of microvesicles (MVs) in promoting self-renewal properties

199 Microvesicles (MVs) released by malignant cancer cells c

200 Our recent evidence indicates that microvesicles (MVs) released by microglia enhance the me

201 Vesicular structures called microvesicles (MVs) that are shed from the surfaces of c

202 L-1b); (2) platelet-derived IL-1b-containing microvesicles (MVs) that increase vascular permeability;

203 cells derived from the ICM generate and shed microvesicles (MVs), a major class of extracellular vesi

204 describe how a specific class of EVs, called microvesicles (MVs), activates VEGF receptors and tumour

205 ere derived from tumour cells, packaged into microvesicles (MVs), and then directly delivered to endo

206 ar vesicles (EVs), specifically exosomes and microvesicles (MVs), are presumed to play key roles in c

207 Microvesicles (MVs), but not exosomes (Exos) or apoptoti

208 lular vesicles (EVs), including exosomes and microvesicles (MVs), by cells has emerged as a form of i

209 llular vesicles, including exosomes and shed microvesicles (MVs), can be internalized by recipient ce

210 18) and their respective fraction carried by microvesicles (MVs), CCL20 and TREM1.

211 lular vesicles (EVs), including exosomes and microvesicles (MVs), have emerged as a major form of int

212 Cell-derived microvesicles (MVs), recognized as important components

213 s phosphatidylserine is a major component of microvesicles (MVs), this study also examined the conseq

214 o smoke extract (TSE) induces the release of microvesicles (MVs; or microparticles) with proteolytic

215 o the distribution of miRNAs among different microvesicles of breast cancer cells, normal cells relea

216 Exosomes are microvesicles of endocytic origin constitutively release

217 hrocyte membrane, with formation of exocytic microvesicles or microparticles and hemolysis, which we

218 cytokines, chemokines, proteases, exosomes, microvesicles, or therapeutic agents, play important and

219 These findings suggest that the microvesicle pathway and P2X7R could represent exploitab

220 As a result, both exosomes and microvesicles play a fundamental biological role in the

221 ndothelial injury and C4d deposition, plasma microvesicles positive for endothelial (CD144) marker an

222 discovered that Tm-CTLA-4 is associated with microvesicles produced by the activated cells.

223 ings indicate that M. tuberculosis increases microvesicle production in response to iron restriction

224 ng that influenza virions form by subverting microvesicle production.

225 Extracellular microvesicles provide local signals (eg, autocrine and p

226 Quantification of plasma C4d+ microvesicles provides information about presence of AMR

227 R on circulating microvesicles, establishing microvesicle release as a novel mechanism for sIL-6R gen

228 T. cruzi metacyclic trypomastigotes induced microvesicle release from blood cells early in infection

229 Notably, firm matrices do not support microvesicle release, whereas compliant matrices are not

230 n of growth factors and their receptors, and microvesicle release.

231 - as a regulator of plasma-membrane-derived microvesicle release.

232 mediated MLC phosphorylation is required for microvesicle release.

233 ubsequent surge in procoagulant activity and microvesicle release.

234 , MSC-derived exosomes (MSC-Exos), a type of microvesicle released from MSCs, were thought to carry f

235 cellular vesicles (EVs) such as exosomes and microvesicles released from cells are potential biomarke

236 Furthermore, we found that TGF-beta-bearing microvesicles released from monocytes and lymphocytes pr

237 esicles, this study examined M. tuberculosis microvesicles released under iron limitation, a common c

238 d plasma membrane origin called exosomes and microvesicles, respectively.

239 n mouse plasma, and infection with exogenous microvesicles resulted in increased T. cruzi parasitemia

240 ecent advances in the study of tumor-derived microvesicles reveal new insights into the cellular basi

241 recently been described as membrane-derived microvesicles secreted by cancer cells, which transfer o

242 the first evidence that cranial grafting of microvesicles secreted from hNSC affords similar neuropr

243 In contrast, their larger cousins, microvesicles, seem to have generally detrimental effect

244 Exosomes and microvesicles share a number of similar characteristics,

245 Microvesicles shed from prostate cancer cells contained

246 endothelial cells through the absorption of microvesicles shed from tumor cells.

247 Microvesicle shedding appears to release selected cellul

248 Microvesicle shedding in tumor cells occurs via an actom

249 To enable microvesicle shedding, ARF6-GTP-dependent activation of

250 on and actomyosin contractility required for microvesicle shedding.

251 d loss of membrane asymmetry associated with microvesicle shedding.

252 ediated phosphorylation of MLC, which blocks microvesicle shedding.

253 racellular vesicles (EVs), exosomes and shed microvesicles (sMVs), which differ in size distribution

254 Our results suggest that tumor microvesicles substantially larger than exosome-sized pa

255 Microvesicles such as exosomes secreted from the iPSC-de

256 determined that exosomes, secreted membrane microvesicles, suppressed the hypoxic pulmonary influx o

257 cantly more B1 receptor-positive endothelial microvesicles than control samples, an effect abrogated

258 -independent cellular process that generates microvesicles that are distinct from exosomes and which,

259 Exosomes are secreted microvesicles that are emerging as potent mediators of c

260 are located on the surface of extracellular microvesicles that bud at the immunological synapse cent

261 Neutrophils deploy extracellular microvesicles that can arrest the growth of bacteria.

262 f endothelial cell-derived microparticles or microvesicles that contain microRNAs which can promote v

263 but also exocytosis, namely of cytokines and microvesicles that contain Nef itself.

264 plasma membrane and mediates the release of microvesicles that contain TSG101, ARRDC1, and other cel

265 stingly, many virally infected cells secrete microvesicles that differ in content from their virion c

266 t T cells release NADPH oxidase 2-containing microvesicles that inhibit TCR activation by elevating R

267 hese mice, releasing mitochondria-containing microvesicles that the epithelia engulfed.

268 Oncosomes are tumor-derived microvesicles that transmit signaling complexes between

269 sonized particles initiated the formation of microvesicles that were able to impair bacterial growth.

270 w cytometry analysis, and significantly more microvesicles that were positive for the kinin B1 recept

271 maturation and shedding of membrane-derived microvesicles, the two key structures involved in invasi

272 In addition, microvesicle tissue factor activity and interleukin-8 le

273 association between plasma interleukin-8 and microvesicle tissue factor activity measured on admissio

274 lation measured on ICU day 1, only increased microvesicle tissue factor activity was significantly as

275 Microvesicle tissue factor activity, thrombin-antithromb

276 aling the connections of microprotrusion and microvesicle to KAI1/CD82 function.

277 gs expand the nucleic acid content of tumour microvesicles to include: elevated levels of specific co

278 ontribution of microRNAs (miRs) delivered by microvesicles to MC activation.

279 Microvesicle treatment was found to attenuate neuroinfla

280 nstrate that pCRP by binding to cell-derived microvesicles undergoes a structural change without disr

281 otic cells was reduced whereas engulfment of microvesicles was increased.

282 e antibacterial effect of neutrophil-derived microvesicles was independent of production of toxic oxy

283 ubjects with AMR, the density of C4d+/CD144+ microvesicles was on average 11-fold (P = 0.002) higher

284 t the exosome fraction of EVs and not larger microvesicles was responsible for induction of TNF-alpha

285 By separating breast cancer cell microvesicles, we find that selectively released miRNAs

286 ties of C4d+ and C4d+/annexin V+ (C4d+/AVB+) microvesicles were also increased in AMR patients compar

287 Neutrophil-derived microvesicles were detected in the serum of healthy dono

288 T cell-derived microvesicles were labeled with PKH67 to allow visualiza

289 olecules contained in the macrophage-derived microvesicles were transported to target cells, includin

290 ene 101 (TSG101) sorts TCRs for inclusion in microvesicles, whereas vacuolar protein sorting 4 (VPS4)

291 Oviductosomes ((OVS), exosomes/microvesicles), which deliver the Ca(2+) efflux pump, pl

292 lular vesicles including structures known as microvesicles, which are known to alter the extracellula

293 s of exosomes, membrane-enclosed subcellular microvesicles, which have immunosuppressive effects on c

294 nelles, exosomes and plasma membrane-derived microvesicles, which were both able to present exogenous

295 ation of microprotrusions and the release of microvesicles, while the mutation disrupts these inhibit

296 the discoveries that (a) they fragment into microvesicles, whose resorption facilitates considerable

297 Macrophages also released microvesicles, whose uptake by epithelial cells was enha

298 review focuses on aspects of the biology of microvesicles with an emphasis on their potential contri

299 solated neutrophilic granulocytes to release microvesicles with different biologic properties.

300 combinant NspA expressed in Escherichia coli microvesicles, with each dose being separated by 3 weeks