The genomic history of the Middle East.

The Middle East region is important to understand human evolution and migrations but is underrepresented in genomic studies. Here, we generated 137 high-coverage physically phased genome sequences from eight Middle Eastern populations using linked-read sequencing. We found no genetic traces of early expansions out-of-Africa in present-day populations but found Arabians have elevated Basal Eurasian ancestry that dilutes their Neanderthal ancestry. Population sizes within the region started diverging 15-20 kya, when Levantines expanded while Arabians maintained smaller populations that derived ancestry from local hunter-gatherers. Arabians suffered a population bottleneck around the aridification of Arabia 6 kya, while Levantines had a distinct bottleneck overlapping the 4.2 kya aridification event. We found an association between movement and admixture of populations in the region and the spread of Semitic languages. Finally, we identify variants that show evidence of selection, including polygenic selection. Our results provide detailed insights into the genomic and selective histories of the Middle East.

Genomic History of Neolithic to Bronze Age Anatolia, Northern Levant, and Southern Caucasus.

Here, we report genome-wide data analyses from 110 ancient Near Eastern individuals spanning the Late Neolithic to Late Bronze Age, a period characterized by intense interregional interactions for the Near East. We find that 6th millennium BCE populations of North/Central Anatolia and the Southern Caucasus shared mixed ancestry on a genetic cline that formed during the Neolithic between Western Anatolia and regions in today's Southern Caucasus/Zagros. During the Late Chalcolithic and/or the Early Bronze Age, more than half of the Northern Levantine gene pool was replaced, while in the rest of Anatolia and the Southern Caucasus, we document genetic continuity with only transient gene flow. Additionally, we reveal a genetically distinct individual within the Late Bronze Age Northern Levant. Overall, our study uncovers multiple scales of population dynamics through time, from extensive admixture during the Neolithic period to long-distance mobility within the globalized societies of the Late Bronze Age. VIDEO ABSTRACT.

Isotopic and DNA analyses reveal multiscale PPNB mobility and migration across Southeastern Anatolia and the Southern Levant.

Growing reliance on animal and plant domestication in the Near East and beyond during the Pre-Pottery Neolithic B (PPNB) (the ninth to eighth millennium BC) has often been associated with a "revolutionary" social transformation from mobility toward more sedentary lifestyles. We are able to yield nuanced insights into the process of the Neolithization in the Near East based on a bioarchaeological approach integrating isotopic and archaeogenetic analyses on the bone remains recovered from Nevali Çori, a site occupied from the early PPNB in Turkey where some of the earliest evidence of animal and plant domestication emerged, and from Ba'ja, a typical late PPNB site in Jordan. In addition, we present the archaeological sequence of Nevali Çori together with newly generated radiocarbon dates. Our results are based on strontium (87Sr/86Sr), carbon, and oxygen (δ18O and δ13Ccarb) isotopic analyses conducted on 28 human and 29 animal individuals from the site of Nevali Çori. 87Sr/86Sr results indicate mobility and connection with the contemporaneous surrounding sites during the earlier PPNB prior to an apparent decline in this mobility at a time of growing reliance on domesticates. Genome-wide data from six human individuals from Nevali Çori and Ba'ja demonstrate a diverse gene pool at Nevali Çori that supports connectedness within the Fertile Crescent during the earlier phases of Neolithization and evidence of consanguineous union in the PPNB Ba'ja and the Iron Age Nevali Çori.

Molecular epidemiology of infectious bursal disease virus in the Near East and Persian Gulf regions.

RESEARCH HIGHLIGHTS: Different field IBDVs were found to circulate in the Near and Middle East.Multiple atypical genotypes (A3B1, A4B1, A6B1) were found to circulate extensively.Traditional very virulent IBDVs (A3B2) were a minority of the detected strains.Viral exchanges can be hypothesized between the region and different continents.

Human and Animal Fascioliasis: Origins and Worldwide Evolving Scenario.

Fascioliasis is a plant- and waterborne zoonotic parasitic disease caused by two trematode species: (i) Fasciola hepatica in Europe, Asia, Africa, the Americas, and Oceania and (ii) F. gigantica, which is restricted to Africa and Asia. Fasciolid liver flukes infect mainly herbivores as ruminants, equids, and camelids but also omnivore mammals as humans and swine and are transmitted by freshwater Lymnaeidae snail vectors. Two phases may be distinguished in fasciolid evolution. The long predomestication period includes the F. gigantica origin in east-southern Africa around the mid-Miocene, the F. hepatica origin in the Near-Middle East of Asia around the latest Miocene to Early Pliocene, and their subsequent local spread. The short postdomestication period includes the worldwide spread by human-guided movements of animals in the last 12,000 years and the more recent transoceanic anthropogenic introductions of F. hepatica into the Americas and Oceania and of F. gigantica into several large islands of the Pacific with ships transporting livestock in the last 500 years. The routes and chronology of the spreading waves followed by both fasciolids into the five continents are redefined on the basis of recently generated knowledge of human-guided movements of domesticated hosts. No local, zonal, or regional situation showing disagreement with historical records was found, although in a few world zones the available knowledge is still insufficient. The anthropogenically accelerated evolution of fasciolids allows us to call them "peridomestic endoparasites." The multidisciplinary implications for crucial aspects of the disease should therefore lead the present baseline update to be taken into account in future research studies.

Giardia duodenalis and dysentery in Iron Age Jerusalem (7th-6th century BCE).

The aim of this study was to determine if the protozoa that cause dysentery might have been present in Jerusalem, the capital of the Kingdom of Judah, during the Iron Age. Sediments from 2 latrines pertaining to this time period were obtained, 1 dating from the 7th century BCE and another from the 7th to early 6th century BCE. Microscopic investigations have previously shown that the users were infected by whipworm (Trichuris trichiura), roundworm (Ascaris lumbricoides), Taenia sp. tapeworm and pinworm (Enterobius vermicularis). However, the protozoa that cause dysentery are fragile and do not survive well in ancient samples in a form recognizable using light microscopy. Enzyme-linked immunosorbent assay kits designed to detect the antigens of Entamoeba histolytica, Cryptosporidium sp. and Giardia duodenalis were used. Results for Entamoeba and Cryptosporidium were negative, while Giardia was positive for both latrine sediments when the analysis was repeated three times. This provides our first microbiological evidence for infective diarrhoeal illnesses that would have affected the populations of the ancient near east. When we integrate descriptions from 2nd and 1st millennium BCE Mesopotamian medical texts, it seems likely that outbreaks of dysentery due to giardiasis may have caused ill health throughout early towns across the region.

Evidence, hints and assumptions for late pregnancy in the Ancient Mediterranean and Near East.

Ancient women, who survived childhood mortality, received good and adequate nutrition, did not work hard and escaped death during childbirth could live fairly long lives. Girls started procreation after marriage, usually at 15 years, had on average seven children, childbearing lasted 14-21 or more years and could happen at the age of 35 or more years. Breastfeeding, usually with contraceptive effect, continued for 2-3 years. Limited actual facts, written evidence and findings, but several hints, assumptions and logical conclusions from secular texts, sacred books, narratives and myths suggest the possibility of late childbearing in the Mediterranean and Near-Eastern ancient world, particularly for the Jews.

Symbolic emblems of the Levantine Aurignacians as a regional entity identifier (Hayonim Cave, Lower Galilee, Israel).

The Levantine Aurignacian is a unique phenomenon in the local Upper Paleolithic sequence, showing greater similarity to the West European classic Aurignacian than to the local Levantine archaeological entities preceding and following it. Herewith we highlight another unique characteristic of this entity, namely, the presence of symbolic objects in the form of notched bones (mostly gazelle scapulae) from the Aurignacian levels of Hayonim Cave, Lower Galilee, Israel. Through both macroscopic and microscopic analyses of the items, we suggest that they are not mere cut marks but rather are intentional (decorative?) human-made markings. The significance of this evidence for symbolic behavior is discussed in its chrono-cultural and geographical contexts. Notched bones are among the oldest symbolic expressions of anatomically modern humans. However, unlike other Paleolithic sites where such findings were reported in single numbers, the number of these items recovered at Hayonim Cave is sufficient to assume they possibly served as an emblem of the Levantine Aurignacian.

Where Asia meets Europe - recent insights from ancient human genomics.

CONTEXT: The peopling of Europe by modern humans is a widely debated topic in the field of modern and ancient genomics. While several recent syntheses have focussed on this topic, little has been discussed about the genetic history of populations in the continent's surrounding regions. OBJECTIVE: We explore genetic transformations in three key areas that played an essential role in the formation of the European genetic landscape through time, focussing on the periods spanning from the Epipalaeolithic/Mesolithic and up until the Iron Age. METHODS: We review published ancient genomic studies and integrate the associated data to provide a quantification and visualisation of major trends in the population histories of the Near East, the western Eurasian Steppe and North East Europe. RESULTS: We describe cross-regional as well as localised prehistoric demographic shifts and discuss potential research directions while highlighting geo-temporal gaps in the data. CONCLUSION: In recent years, archaeogenetic studies have contributed to the understanding of human genetic diversity through time in regions located at the doorstep of Europe. Further studies focussing on these areas will allow for a better characterisation of genetic shifts and regionally-specific patterns of admixture across western Eurasia.

Continuity and Admixture in the Last Five Millennia of Levantine History from Ancient Canaanite and Present-Day Lebanese Genome Sequences.

The Canaanites inhabited the Levant region during the Bronze Age and established a culture that became influential in the Near East and beyond. However, the Canaanites, unlike most other ancient Near Easterners of this period, left few surviving textual records and thus their origin and relationship to ancient and present-day populations remain unclear. In this study, we sequenced five whole genomes from ∼3,700-year-old individuals from the city of Sidon, a major Canaanite city-state on the Eastern Mediterranean coast. We also sequenced the genomes of 99 individuals from present-day Lebanon to catalog modern Levantine genetic diversity. We find that a Bronze Age Canaanite-related ancestry was widespread in the region, shared among urban populations inhabiting the coast (Sidon) and inland populations (Jordan) who likely lived in farming societies or were pastoral nomads. This Canaanite-related ancestry derived from mixture between local Neolithic populations and eastern migrants genetically related to Chalcolithic Iranians. We estimate, using linkage-disequilibrium decay patterns, that admixture occurred 6,600-3,550 years ago, coinciding with recorded massive population movements in Mesopotamia during the mid-Holocene. We show that present-day Lebanese derive most of their ancestry from a Canaanite-related population, which therefore implies substantial genetic continuity in the Levant since at least the Bronze Age. In addition, we find Eurasian ancestry in the Lebanese not present in Bronze Age or earlier Levantines. We estimate that this Eurasian ancestry arrived in the Levant around 3,750-2,170 years ago during a period of successive conquests by distant populations.

Status, kinship, and place of burial at Early Bronze Age Bab adh-Dhra': A biogeochemical comparison of charnel house human remains.

OBJECTIVES: The Early Bronze Age (EBA; ca. 3,600-2000 BCE) of the southern Levant underwent considerable transformation as agro-pastoral communities began to utilize their land more intensively, constructing larger, fortified towns prior to site abandonment at the end of the third millennium. At the site of Bab adh-Dhra' in Jordan, the dead of the Early Bronze (EB) II-III (ca. 3,100-2,500 BCE) period were communally interred within charnel houses, but important disparities between these structures and their contents may be reflective of ownership and use by particular extended kin groups whose activity patterns, subsistence strategies, and even social status may have differed from one another. Subsequently, we hypothesized that differences in mobility and dietary intake may differentiate tomb groups from one another. MATERIALS AND METHODS: Dental enamel from 31 individuals interred in three different Early Bronze Age charnel houses (A56, A22, A55) at Bab adh-Dhra', Jordan were analyzed for strontium, oxygen, and carbon isotope values. RESULTS: Strontium isotope ratios (range: 0.70793-0.70842) possessed medians that did not differ statistically from one another, but had ranges that exhibited significant differences in variance. Carbon isotope values ( x¯ = -13.2 ± 0.5‰, 1σ) were not significantly different. DISCUSSION: General similarities in human isotopic signatures between EB II-III charnel houses A22 and A55 suggest that their activities were likely similar to one another and agree with findings from excavated domestic spaces with little archaeological evidence for economic, social, or political differentiation. More variable strontium isotope ratios and lower carbon isotope values from A22 could reflect a greater involvement with pastoralist practices or regional trade, including the consumption of more 13 C-depleted foods, while those in A55 may have led a more sedentary lifestyle with greater involvement in cultivating orchard crops. All charnel houses contained nonlocal individuals likely originating from other Dead Sea Plain sites with no EB II-III cemeteries of their own, supporting the idea that extended kin groups throughout the region returned to Bab adh-Dhra' to bury their dead.

Early Neolithic wine of Georgia in the South Caucasus.

Chemical analyses of ancient organic compounds absorbed into the pottery fabrics from sites in Georgia in the South Caucasus region, dating to the early Neolithic period (ca. 6,000-5,000 BC), provide the earliest biomolecular archaeological evidence for grape wine and viniculture from the Near East, at ca. 6,000-5,800 BC. The chemical findings are corroborated by climatic and environmental reconstruction, together with archaeobotanical evidence, including grape pollen, starch, and epidermal remains associated with a jar of similar type and date. The very large-capacity jars, some of the earliest pottery made in the Near East, probably served as combination fermentation, aging, and serving vessels. They are the most numerous pottery type at many sites comprising the so-called "Shulaveri-Shomutepe Culture" of the Neolithic period, which extends into western Azerbaijan and northern Armenia. The discovery of early sixth millennium BC grape wine in this region is crucial to the later history of wine in Europe and the rest of the world.

Analyses of Neanderthal introgression suggest that Levantine and southern Arabian populations have a shared population history.

OBJECTIVES: Modern humans are thought to have interbred with Neanderthals in the Near East soon after modern humans dispersed out of Africa. This introgression event likely took place in either the Levant or southern Arabia depending on the dispersal route out of Africa that was followed. In this study, we compare Neanderthal introgression in contemporary Levantine and southern Arabian populations to investigate Neanderthal introgression and to study Near Eastern population history. MATERIALS AND METHODS: We analyzed genotyping data on >400,000 autosomal SNPs from seven Levantine and five southern Arabian populations and compared these data to those from populations from around the world including Neanderthal and Denisovan genomes. We used f4 and D statistics to estimate and compare levels of Neanderthal introgression between Levantine, southern Arabian, and comparative global populations. We also identified 1,581 putative Neanderthal-introgressed SNPs within our dataset and analyzed their allele frequencies as a means to compare introgression patterns in Levantine and southern Arabian genomes. RESULTS: We find that Levantine and southern Arabian populations have similar levels of Neanderthal introgression to each other but lower levels than other non-Africans. Furthermore, we find that introgressed SNPs have very similar allele frequencies in the Levant and southern Arabia, which indicates that Neanderthal introgression is similarly distributed in Levantine and southern Arabian genomes. DISCUSSION: We infer that the ancestors of contemporary Levantine and southern Arabian populations received Neanderthal introgression prior to separating from each other and that there has been extensive gene flow between these populations.

New chronology for Ksâr 'Akil (Lebanon) supports Levantine route of modern human dispersal into Europe.

Modern human dispersal into Europe is thought to have occurred with the start of the Upper Paleolithic around 50,000-40,000 y ago. The Levantine corridor hypothesis suggests that modern humans from Africa spread into Europe via the Levant. Ksâr 'Akil (Lebanon), with its deeply stratified Initial (IUP) and Early (EUP) Upper Paleolithic sequence containing modern human remains, has played an important part in the debate. The latest chronology for the site, based on AMS radiocarbon dates of shell ornaments, suggests that the appearance of the Levantine IUP is later than the start of the first Upper Paleolithic in Europe, thus questioning the Levantine corridor hypothesis. Here we report a series of AMS radiocarbon dates on the marine gastropod Phorcus turbinatus associated with modern human remains and IUP and EUP stone tools from Ksâr 'Akil. Our results, supported by an evaluation of individual sample integrity, place the EUP layer containing the skeleton known as "Egbert" between 43,200 and 42,900 cal B.P. and the IUP-associated modern human maxilla known as "Ethelruda" before ∼ 45,900 cal B.P. This chronology is in line with those of other Levantine IUP and EUP sites and demonstrates that the presence of modern humans associated with Upper Paleolithic toolkits in the Levant predates all modern human fossils from Europe. The age of the IUP-associated Ethelruda fossil is significant for the spread of modern humans carrying the IUP into Europe and suggests a rapid initial colonization of Europe by our species.

The Multiple Histories of Western Asia: Perspectives from Ancient and Modern Genomes.

Western Asia lies at the heart of the Old World, in the midst of Africa, Asia, and Europe. As such, this region has been populated and repopulated by myriad peoples, starting with the first migrants from Africa. All evidence points to Western Asia for the beginnings of sedentary life, and indeed, first the villages and later the cities of this land remain as archaeological wonders, revealing complex histories of multiple peoples and their interactions. With the wondrous breakthroughs in genomic studies, we now have the power to look at these histories with a truly quantitative lens. Here, we review the recent anthropological genomics literature pertaining to this region, with an outlook for the future challenges and exciting possibilities for the field.

Phylogeny and diversification of mountain vipers (Montivipera, Nilson et al., 2001) triggered by multiple Plio-Pleistocene refugia and high-mountain topography in the Near and Middle East.

The Near and Middle East is a hotspot of biodiversity, but the region remains underexplored at the level of genetic biodiversity. Here, we present an extensive molecular phylogeny of the viperid snake genus Montivipera, including all known taxa. Based on nuclear and mitochondrial data, we present novel insights into the phylogeny of the genus and review the status of its constituent species. Maximum likelihood methods revealed a montane origin of Montivipera at 12.3Mya. We then analyzed factors of mountain viper diversity. Our data support substantial changes in effective population size through Plio-Pleistocene periods. We conclude that climatic oscillations were drivers of allopatric speciation, and that mountain systems of the Near and Middle East have strongly influenced the evolution and survival of taxa, because climatic and topographical heterogeneities induced by mountains have played a crucial role as filters for dispersal and as multiple refugia. The wide diversity of montane microhabitats enabled mountain vipers to retain their ecological niche during climatic pessima. In consequence the varied geological and topographical conditions between refugia favoured genetic isolation and created patterns of species richness resulting in the formation of neoendemic taxa. Our data support high concordance between geographic distributions of Montivipera haplotypes with putative plant refugia.

Monks on the move: Evaluating pilgrimage to Byzantine St. Stephen's monastery using strontium isotopes.

OBJECTIVES: Written and archaeological evidence indicates that migration, in the form of pilgrimage to Byzantine Palestine, was a major social phenomenon between the 5th and 7th centuries CE. Monasteries saw record growth at this time as pilgrims chose to stay in the region and take up religious life. A major influx of people to the region, with a corresponding growth in monastic vocations that led travelers to stay in the area, is not in question; however, the distant origins of pilgrims reflected in surviving texts may be an artifact of preservation, biased towards an elite minority. The Byzantine monastery of St. Stephen's provides an opportunity to study this question from a bioarchaeological perspective, given the excellent preservation of the human skeletal assemblage, a wealth of written works about the community as well as daily life in Byzantine Palestine, and a rich archaeological record for the site and region. MATERIALS AND METHODS: An analysis of radiogenic strontium isotope values from the third molars of 22 individuals recovered from the St. Stephen's crypt complex was conducted to test whether those interred at the monastery were of local origin. RESULTS: Of those examined ( x¯= 0.7084 ± 0.0007, 1σ), 8 out of 22 (36%) exhibited (87) Sr/(86) Sr ratios that fell outside of local ranges. DISCUSSION: These results confirmed the sizeable presence of nonlocals at St. Stephen's Monastery. While most of these migrants likely traveled to Jerusalem from different areas of the Levant as pilgrims, others may have hailed from further afield, including Europe.

The prevalence of pulp stones in historical populations from the middle Euphrates valley (Syria).

There are many reports in the literature concerning pulp stones in contemporary living populations, but there are no descriptions of cases of pulp stones and their prevalence in populations of the past. Here we present a study of pulp stones in a series of archaeologically derived samples from the Middle Euphrates Valley (Syria) obtained from two sites: Terqa and Tell Masaikh. The specimens were assigned to five periods: Early Bronze (2650-2350 BC); Middle Bronze (2200-1700 BC); late Roman (AD 200-400); Islamic (AD 600-1200); and Modern Islamic (AD 1850-1950). A total of 529 teeth representing 117 adult individuals of both sexes were examined. Pulp stones were identified by X-ray and 10 selected specimens were sectioned for histological study. Pulp stones were found in 99 of 117 individuals (85%) and in 271 of 529 (51%) teeth. Pulp stone prevalence was found to increase with age, for individuals of older age classes have more pulp stones than younger individuals. Intriguingly, the prevalence of single pulp stones was higher among older individuals (36-45, >46), while younger individuals (17-25, 26-35) more often possessed multiple stones. Individuals with moderate to highly advanced dental wear have pulp stones significantly more often than individuals whose tooth wear is limited to invisible or very small facets. Though there is no statistical significance in the prevalence of pulp stones across chronological periods, it appears that a high level of calcium in the diet is accompanied by a greater prevalence of pulp stones.

Temporal pattern of dental caries at the western flank of the Central Plateau of Iran, c. 2700 BCE - 1600 CE.

OBJECTIVE: To analyze the overall frequency and inter-tooth patterns of caries in three populations from ancient cemeteries located along the western border of the Central Iranian Plateau as a means to explore whether the populations of Iran had greater access to fermentable sugars after the establishment of the great empires. MATERIALS: Dental collections from Kafarved-Varzaneh (Early Bronze Age, MNI=66), Estark-Joshaqan (Iron Age, MNI=57), Tappeh Poustchi (Timurid and Safavid Period, MNI=34), together with comparative data from NE Syria. METHODS: Frequencies of dental caries per tooth categories, location and size of carious lesions are analyzed using Smith's Mean Measure of Divergence, Correspondence Analysis, χ2 and Kruskal-Wallis tests. RESULTS: There are minimal differences in overall frequencies of carious lesions at Iranian sites, regardless of the chronology, but notable differences at Syrian sites. The inter-tooth pattern at the Iron Age cemetery in Estark appears distinctly different than the other Iranian sites and the comparative samples from Syria. CONCLUSIONS: Divergent subsistence strategies may be linked with different inter-tooth patterns since people buried at Estark were mobile herders, while the other cemeteries were used by settled farmers. SIGNIFICANCE: This comprehensive research on dental caries in three chronologically diverse populations in Iran sheds light on the association between dental caries and subsistence strategies, and introduces the Smith's Mean Measure of Divergence to explore inter-tooth carious patterns, which may prove useful to other researchers seeking to understand the relationships between subsistence, diet, and the presence of carious lesions. LIMITATIONS: The studied sample size is relatively small and therefore its temporal/regional distribution produces low-resolution results. SUGGESTIONS FOR FURTHER RESEARCH: More systematic research on the patterns of dental caries is necessary to produce more fine-grained reconstructions of diet and subsistence in Iran and around the globe.

Antiproliferative and immunomodulative potential of Citrullus colocynthis and its bioactive compounds in human lymphocytes and lung cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Citrullus colocynthis (L.) Schrad is a member of the Cucurbitaceae plant family which has been used in traditional medicine for the treatment of lung diseases such as asthma and bronchitis. AIM OF THE STUDY: The study was conducted to investigate antiproliferative and immunomodulating effects of C. colocynthis and isolated cucurbitacins on human T lymphocytes and lung epithelial cells in order to evaluate their potential in the treatment of airway diseases. MATERIALS AND METHODS: Different concentrations of an ethanolic extract of C. colocynthis fruits and cucurbitacins B (CuB), E (CuE) and E-glucopyranoside (CuE-Glu) were analysed for their cytotoxicity and immunomodulatory potential on Peripheral Blood Mononuclear Cells (PBMCs) of healthy donors and on the epithelial lung cancer cell line A549. Viability and proliferation were tested using WST1 and CFSE assays. Flow cytometric analysis of AnnexinV/PI staining was used to investigate cell death through apoptosis/necrosis. Effects on regulatory mechanisms of T lymphocytes, such as CD69 and CD25 marker activation, cytokine production of the cytokines interleukin 2 (IL2), tumor necrosis factor α (TNFα) and interferon γ (IFNy) were also analysed via flow cytometry. Influences on the activator protein 1 (AP1), nuclear factor of activated T-cells (NFAT) or nuclear factor 'kappa-light-chain-enhancer' of activated B-cells (NFκB) pathways were analysed in the Jurkat reporter cell line. Cytokine secretion in A549 cells stimulated with virus-like particles was analysed using the bead-based Legendplex™ assay. RESULTS: Non-toxic concentrations of C. colocynthis and CuE-Glu showed dose-dependent effects on viability and proliferation in both T lymphocytes and A549 cells. The extracts inhibited lymphocyte activation and suppressed T cell effector functions, which was also shown by lower production of cytokines IL2, TNFα and IFNy. A dose dependent inhibition of the pathways NFκB, NFAT and AP1 in Jurkat cells could be observed. In A549 cells, especially CuE and CuE-Glu showed inhibitory effects on cytokine production following a simulated viral infection. Unglycosylated cucurbitacins were more effective in suppressing the immune function in lymphocytes than glycosylated cucurbitacins, however this activity is limited to cytotoxic concentrations. CONCLUSION: In our study we could confirm the immunmodulating effect of C. colocynthis and cucurbitacins B, E and E-glucopyranoside in vitro by suppression of different pathways of inflammation and T cell proliferation. Activity in a lung cell model using a virus-like stimulation shows promise for further research regarding cucurbitacins in airway diseases.